Ortho-substituted benzoic acid derivatives for the treatmetn of insulin resistance

ABSTRACT

The present invention provides a compound of formula (I), wherein n is 0, 1 or 2 and R 1  represents halo, a C 1-4 alkyl group which is optionally substituted by one or more fluoro, a C 1-4 alkoxy group which is optionally substituted by one or more fluoro and wherein when n is 2 the substituents R 1  may be the same or different; R 2  represents a C 2-8 alkyl group which is optionally interrupted by oxygen; Y is absent or represents methylene; and X is O or S; and pharmaceutically acceptable salts and prodrugs thereof, to processes for preparing such compounds, to their utility in treating clinical conditions associated with insulin resistance, to methods for their therapeutic use and to pharmaceutical compositions containing them.

FIELD OF THE INVENTION

The present invention relates to certain novel benzoic acid derivatives,to processes for preparing such compounds, to their utility in treatingclinical conditions associated with insulin resistance, to methods fortheir therapeutic use and to pharmaceutical compositions containingthem.

BACKGROUND OF THE INVENTION

The Insulin Resistance Syndrome (IRS) including type 2 diabetesmellitus, which refers to a cluster of manifestations including insulinresistance with accompanying hyperinsulinaemia, possible type 2 diabetesmellitus, arterial hypertension, central (visceral) obesity,dyslipidaemia observed as deranged lipoprotein levels typically ischaracterised by elevated VLDL (very low density lipoproteins), smalldense LDL particles and reduced HDL (high density lipoprotein)concentrations and reduced fibrinolysis.

Recent epidemiological research has documented that individuals withinsulin resistance run a greatly increased risk of cardiovascularmorbidity and mortality, notably suffering from myocardial infarctionand stroke. In type 2 diabetes mellitus atherosclerosis relatedconditions cause up to 80% of all deaths.

In clinical medicine there is awareness of the need to increase theinsulin sensitivity in IRS suffering patients and thus to correct thedyslipidaemia which is considered to cause the accelerated progress ofatherosclerosis. However, currently this is not a universally welldefined disease.

Modulators of peroxisome proliferator-activated receptors (PPAR, for areview of the PPARs see T. M. Willson et al, J Med Chem 2000, Vol 43,527) are effective in treating conditions associated with insulinresistance.

U.S. Pat. No. 5,750,783 discloses that certain benzyloxy-substitutedphenylglycinolamides which have a cycloalkyl substituent areantiatherosclerotic medicaments. This document does not disclose orsuggest the compounds of the present invention. Surprisingly a series ofcompounds has now been found which are selective PPARα modulators.

DESCRIPTION OF THE INVENTION

The present invention provides a compound of formula I

wherein n is 0, 1 or 2 and R¹ represents halo, a C₁₋₄alkyl group whichis optionally substituted by one or more fluoro, a C₁₋₄alkoxy groupwhich is optionally substituted by one or more fluoro and wherein when nis 2 the substituents R¹ may be the same or different;

-   R² represents a C₂₋₈alkyl group which is optionally interrupted by    oxygen;-   Y is absent or represents methylene; and-   X is O or S;    and pharmaceutically acceptable salts and prodrugs thereof.

Further values of R¹, R², Y and X in compounds of Formula I now follow.It will be understood that such values may be used where appropriatewith any of the definitions, claims or embodiments defined hereinbeforeor hereinafter.

-   In one aspect X is O.-   In a second aspect X is S.-   In a third aspect Y is methylene.-   In a fourth aspect Y is absent.-   In a fifth aspect R¹ is halo, a C₁₋₄alkyl group or a C₁₋₄alkoxy    group and n is 0, 1 or 2.-   Particularly R¹ is fluoro, methoxy, or isopropyl when n is 1 or 2.    Particularly n is 0.-   In a sixth aspect R² represents a C₅₋₇alkyl group.

The term C₂₋₈alkyl denotes a straight-chain or branched saturatedaliphatic hydrocarbon having from 2 to 8 carbon atoms. Examples of saidalkyl include ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl,sec-butyl, t-butyl and straight- and branched-chain pentyl, hexyl,heptyl and octyl.

It will be understood by those skilled in the art that the terminterrupted as used above means that the oxygen atom is situated withinthe alkyl chain and is not the terminal atom. The term “prodrug” as usedin this specification includes derivatives of the carboxylic acid groupwhich are converted in a mammal, particularly a human, into thecarboxylic acid group or a salt or conjugate thereof. It should beunderstood that, whilst not being bound by theory, it is believed thatmost of the activity associated with the prodrugs arises from theactivity of the compound of formula I into which the prodrugs areconverted. Prodrugs can be prepared by routine methodology well withinthe capabilities of someone skilled in the art. Various prodrugs ofcarboxy are known in the art. For examples of such prodrug derivatives,see:

-   a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) and    Methods in Enzymology. 42: 309-396, edited by K. Widder, et al.    (Academic Press, 1985);-   b) A Textbook of Drug Design and Development, edited by    Krogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design and    Application of Prodrugs”, by H. Bundgaard p. 113-191 (1991);-   c) H. Bundgaard, Advanced Drug Delivery Reviews, 8:1-38 (1992);-   d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77:285    (1988); and-   e) N. Kalceya, et al., Chem Pharm Bull, 32:692 (1984).    The above documents a to e are herein incorporated by reference.    In vivo cleavable esters are just one type of prodrug of the parent    molecule.

The compounds of formula I have activity as medicaments, in particularthe compounds of formula I are selective agonists of PPARα, that is,their EC₅₀ for PPARα is at least three times lower, preferably at leastfour times lower and more preferably 10 or 50 times lower than theirrespective EC₅₀ for PPARγ wherein the EC₅₀s are measured and calculatedas described in the assays later in this document. The compounds offormula I are potent and selective.

Specific compounds of the invention are one or more of the following:

-   2-[(4-{3-[benzyl(hexyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;-   2-{[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoic    acid;-   2-[(4-{2-[benzyl(hexyl)amino]-2-oxoethyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;-   2-{[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenyl)thio]methyl}benzoic    acid;-   2-[(4-{3-[butyl(2,3-dimethoxybenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{3-[(2,3-dimethoxybenzyl)(heptyl)-amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{3-[(3-ethoxypropyl)(4-isopropylbenzyl)amino]-3-oxopropyl}phenoxy)methyl]-benzoic    acid;-   2-[(4-{3-[(2,4-difluorobenzyl)(propyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{2-[ethyl(2-fluorobenzyl)amino]-2-oxoethyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{3-[ethyl(2-fluorobenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;-   2-({[4-(2-{ethyl[4-(trifluoromethyl)benzyl]amino}-2-oxoethyl)phenyl]thio}-methyl)benzoic    acid;-   2-{[(3-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoic    acid; and-   2-{[(4-{2-[(4-chlorobenzyl)(ethyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoic    acid and pharmaceutically acceptable salts thereof.

A second group of specific compounds of the invention comprises acompound selected from:

-   2-{[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoic    acid;-   2-{[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenyl)thio]methyl}benzoic    acid;-   2-({[4-(2-{ethyl[4-(trifluoromethyl)benzyl]amino}-2-oxoethyl)phenyl]thio}-methyl)benzoic    acid;-   2-{[(3-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoic    acid; and-   2-{[(4-{2-[(4-chlorobenzyl)(ethyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoic    acid and pharmaceutically acceptable salts thereof.

A third group of specific compounds of the invention comprises acompound selected from:

-   2-[(4-{3-[benzyl(hexyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{2-[benzyl(hexyl)amino]-2-oxoethyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{3-[butyl(2,3-dimethoxybenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{3-[(2,3-dimethoxybenzyl)(heptyl)-amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{3-[(3-ethoxypropyl)(4-isopropylbenzyl)amino]-3-oxopropyl}phenoxy)methyl]-benzoic    acid;-   2-[(4-{3-[(2,4-difluorobenzyl)(propyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{2-[ethyl(2-fluorobenzyl)amino]-2-oxoethyl}phenoxy)methyl]benzoic    acid;-   2-[(4-{3-[ethyl(2-fluorobenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic    acid;    and pharmaceutically acceptable salts thereof.

Certain compounds of the present invention may exist as tautomers. It isto be understood that the present invention encompasses all suchtautomers.

Throughout the specification and the appended claims, a given chemicalformula or name shall encompass all stereo and optical isomers andracemates thereof as well as mixtures in different proportions of theseparate enantiomers, where such isomers and enantiomers exist, as wellas pharmaceutically acceptable salts thereof. Isomers may be separatedusing conventional techniques, e.g. chromatography or fractionalcrystallisation. The enantiomers may be isolated by separation ofracemate for example by fractional crystallisation, resolution or HPLC.The diastereomers may be isolated by separation of isomer mixtures forinstance by fractional crystallisation, HPLC or flash chromatography.Alternatively the stereoisomers may be made by chiral synthesis fromchiral starting materials under conditions which will not causeracemisation or epimerisation, or by derivatisation, with a chiralreagent. All stereoisomers are included within the scope of theinvention.

Methods of Preparation

The compounds of the invention may be prepared as outlined below.However, the invention is not limited to these methods, the compoundsmay also be prepared as described for structurally related compounds inthe prior art. The reactions can be carried out according to standardprocedures or as described in the experimental section.

Compounds of formula I may be prepared by reacting a compound of formulaII

in which R¹, R², X and Y are as previously defined and PG represents aprotecting group for a carboxylic hydroxy group as described in thestandard text “Protective Groups in Organic Synthesis”, 2^(nd) Edition(1991) by Greene and Wuts, with a de-protecting agent.

The protecting group may also be a resin, such as Wang resin or2-chlorotrityl chloride resin. Protecting groups may be removed inaccordance to techniques which are well known to those skilled in theart. One such protecting group is where PG represents a C₁₋₆alkoxy groupor an arylalkoxy group eg benzyl, such that COPG represents an ester.Such esters can be reacted with a hydrolysing agent, for example lithiumhydroxide in the presence of a solvent for example a mixture of THF andwater or potassium hydroxide in a C₁₋₃ alcohol for example methanol, ata temperature in the range of 0-200° C. or by microwave radiation togive compounds of formula I.

Compounds of formula II may be prepared by reacting a compound offormula III

or a salt thereof, for example a hydrochloride salt, in which R¹, R² andn are as previously defined with a compound of formula IV

or the acid chloride thereof in which X, Y and PG are as previouslydefined in an inert solvent, for example dichloromethane, optionally inthe presence of a coupling agent, for example 4-dimethylaminopyridine or1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, at atemperature in the range of −25° C. to 150° C.

Compounds of formula II may also be prepared by reacting a compound offormula V

in which R¹, n, R², X and Y are as previously defined with a compound offormula VI

in which PG is as previously defined and L represents a leaving group,for example halo, is e.g. bromo, optionally in the presence of solvent,for example acetonitrilie, and optionally in the presence of a base, forexample potassium carbonate, at a temperature in the range of 0 to 150°C.

Compounds of formula m, IV, V and VI may be prepared by methodsdescribed in the Examples or by analogous methods known to those skilledin the art.

Compounds of formula II, III, IV and V are useful intermediates in thepreparation of compounds of formula I. Certain of these compounds arebelieved to be novel. Novel compounds of formula II, or formula III, orformula IV or formula V are herein claimed as a further aspect of thepresent invention.

The compounds of the invention may be isolated from their reactionmixtures using conventional techniques.

Persons skilled in the art will appreciate that, in order to obtaincompounds of the invention in an alternative and in some occasions, moreconvenient manner, the individual process steps mentioned hereinbeforemay be performed in different order, and/or the individual reactions maybe performed at different stage in the overall route (i.e. chemicaltransformations may be performed upon different intermediates to thoseassociated hereinbefore with a particular reaction).

In any of the preceding methods of preparation, where necessary,hydroxy, amino or other reactive groups may be protected using aprotecting group, R^(p) as described in the standard text “Protectivegroups in Organic Synthesis”, 2^(nd) Edition (1991) by Greene and Wuts.The protecting group may also be a resin, such as Wang resin or2-chlorotrityl chloride resin. The protection and deprotection offunctional groups may take place before or after any of the reactionsteps described hereinbefore. Protecting groups may be removed inaccordance to techniques which are well known to those skilled in theart.

The expression “inert solvent” refers to a solvent which does not reactwith the starting materials, reagents, intermediates or products in amanner which adversely affects the yield of the desired product.

Pharmaceutical Preparations

The compounds of the invention will normally be administered via theoral, parenteral, intravenous, intramuscular, subcutaneous or in otherinjectable ways, buccal, rectal, vaginal, transdermal and/or nasal routeand/or via inhalation, in the form of pharmaceutical preparationscomprising the active ingredient either as a free acid or as apharmaceutically acceptable salt, in a pharmaceutically acceptabledosage form. Depending upon the disorder and patient to be treated andthe route of administration, the compositions may be administered atvarying doses.

Suitable daily doses of the compounds of the invention in therapeuticaltreatment of humans are about 0.0001-100 mg/kg body weight, preferably0.001-10 mg/kg body weight.

Oral formulations are preferred particularly tablets or capsules whichmay be formulated by methods known to those skilled in the art toprovide doses of the active compound in the range of 0.5 mg to 500 mgfor example 1 mg, 3 mg, 5 mg, 10 mg, 25 mg, 50 mg, 100 mg and 250 mg.

According to a further aspect of the invention there is thus provided apharmaceutical formulation including any of the compounds of theinvention, or a pharmaceutically acceptable salt thereof, in admixturewith pharmaceutically acceptable adjuvants, diluents and/or carriers.

Pharmacological Properties

The present compounds of formula (I) are useful for the prophylaxisand/or treatment of clinical conditions associated with inherent orinduced reduced sensitivity to insulin (insulin resistance) andassociated metabolic disorders (also known as metabolic syndrome). Theseclinical conditions will include, but will not be limited to, generalobesity, abdominal obesity, arterial hypertension, hyperinsulinaemia,hyperglycaemia, type 2 diabetes and the dyslipidaemia characteristicallyappearing with insulin resistance. This dyslipidaemia, also known as theatherogenic lipoprotein profile, is characterised by moderately elevatednon-esterified fatty acids, elevated very low density lipoprotein (VLDL)triglyceride rich particles, high Apo B levels, low high densitylipoprotein (ML) levels associated with low apoAI particle levels andhigh Apo B levels in the presence of small, dense, low densitylipoproteins (LDL) particles, phenotype B.

The compounds of the present invention are expected to be useful intreating patients with combined or mixed hyperlipidemias or variousdegrees of hypertriglyceridemias and postprandial dyslipidemia with orwithout other manifestations of the metabolic syndrome.

Treatment with the present compounds is expected to lower thecardiovascular morbidity and mortality associated with atherosclerosisdue to their antidyslipidaemic as well as antiinflammatory properties.The cardiovascular disease conditions include macro-angiopathies ofvarious internal organs causing myocardial infarction, congestive heartfailure, cerebrovascular disease and peripheral arterial insufficiencyof the lower extremities. Because of their insulin sensitizing effectthe compounds of formula I are also expected to prevent or delay thedevelopment of type 2 diabetes from the metabolic syndrome and diabetesof pregnancy. Therefore the development of long-term complicationsassociated with chronic hyperglycaemia in diabetes mellitus such as themicro-angiopathies causing renal disease, retinal damage and peripheralvascular disease of the lower limbs are expected to be delayed.Furthermore the compounds may be useful in treatment of variousconditions outside the cardiovascular system whether or not associatedwith insulin resistance, like polycystic ovarian syndrome, obesity,cancer and states of inflammatory disease including neurodegenerativedisorders such as mild cognitive impairment, Alzheimer's disease,Parkinson's disease and multiple sclerosis.

The compounds of the present invention are expected to be useful incontrolling glucose levels in patients suffering from type 2 diabetes.

The present invention provides a method of treating or preventingdyslipidemias, the insulin resistance syndrome and/or metabolicdisorders (as defined above) comprising the administration of a compoundof formula I to a mammal (particularly a human) in need thereof.

The present invention provides a method of treating or preventing type 2diabetes comprising the administration of an effective amount of acompound of formula I to a mammal (particularly a human) in needthereof.

In a further aspect the present invention provides the use of a compoundof formula I as a medicament.

In a further aspect the present invention provides the use of a compoundof formula I in the manufacture of a medicament for the treatment ofinsulin resistance and/or metabolic disorders.

Combination Therapy

The compounds of the invention may be combined with another therapeuticagent that is useful in the treatment of disorders associated with thedevelopment and progress of atherosclerosis such as hypertension,hyperlipidaemias, dyslipidaemias, diabetes and obesity. The compounds ofthe invention may be combined with another therapeutic agent thatdecreases the ratio of LDL:HDL or an agent that causes a decrease incirculating levels of LDL-cholesterol. In patients with diabetesmellitus the compounds of the invention may also be combined withtherapeutic agents used to treat complications related tomicro-angiopathies. The compounds of the invention may be used alongsideother therapies for the treatment of metabolic syndrome or type 2diabetes and its associated complications, these include biguanidedrugs, for example metformin, phenformin and buformin, insulin(synthetic insulin analogues, amylin) and oral antihyperglycemics (theseare divided into prandial glucose regulators and alpha-glucosidaseinhibitors). An example of an alpha-glucosidase inhibitor is acarbose orvoglibose or miglitol. An example of a prandial glucose regulator isrepaglinide or nateglinide.

In another aspect of the invention, the compound of formula I, or apharmaceutically acceptable salt, solvate, solvate of such a salt or aprodrug thereof, may be administered in association with another PPARmodulating agent. PPAR modulating agents include but are not limited toa PPAR alpha and/or gamma and/or delta agonist, or pharmaceuticallyacceptable salts, solvates, solvates of such salts or prodrugs thereof.Suitable PPAR alpha and/or gamma agonists, pharmaceutically acceptablesalts, solvates, solvates of such salts or prodrugs thereof are wellknown in the art. These include the compounds described in WO 01/12187,WO 01/12612, WO 99/62870, WO 99/62872, WO 99/62871, WO 98/57941, WO01/40170, J Med Chem, 1996, 39, 665, Expert Opinion on TherapeuticPatents, 10 (5), 623-634 (in particular the compounds described in thepatent applications listed on page 634) and J Med Chem, 2000, 43, 527which are all incorporated herein by reference. Particularly a PPARalpha and/or gamma agonist refers to BMS 298585, clofibrate,fenofibrate, bezafibrate, gemfibrozil and ciprofibrate; GW 9578,pioglitazone, rosiglitazone, rivoglitazone, balaglitazone, KRP-297,JTT-501, SB 213068, GW 1929, GW 7845, GW 0207, L-796449, L-165041 and GW2433. Particularly a PPAR alpha and/or gamma agonist refers to(S)-2-ethoxy-3-[4-(2-{4-methanesulphonyloxy-phenyl}ethoxy)phenyl]propanoicacid and pharmaceutically acceptable salts thereof.

In addition the combination of the invention may be used in conjunctionwith a sulfonylurea for example: glimepiride, glibenclamide (glyburide),gliclazide, glipizide, gliquidone, chloropropamide, tolbutamide,acetohexamide, glycopyramide, carbutamide, glibonuride, glisoxepid,glybuthiazole, glibuzole, glyhexamide, glymidine, glypinamide,phenbutamide, tolcylamide and tolazamide. Preferably the sulfonylurea isglimepiride or glibenclamide (glyburide). More preferably thesulfonylurea is glimepiride. Therefore the present invention includesadministration of a compound of the present invention in conjunctionwith one, two or more existing therapies described in this paragraph.The doses of the other existing therapies for the treatment of type 2diabetes and its associated complications will be those known in the artand approved for use by regulatory bodies for example the FDA and may befound in the Orange Book published by the FDA. Alternatively smallerdoses may be used as a result of the benefits derived from thecombination. The present invention also includes a compound of thepresent invention in combination with a cholesterol-lowering agent. Thecholesterol-lowering agents referred to in this application include butare not limited to inhibitors of HMG-CoA reductase(3-hydroxy-3-methylglutaryl coenzyme A reductase). Suitably the HMG-CoAreductase inhibitor is a statin selected from the group consisting ofatorvastatin, bervastatin, cerivastatin, dalvastatin, fluvastatin,itavastatin, lovastatin, mevastatin, nicostatin, nivastatin, pravastatinand simvastatin, or a pharmaceutically acceptable salt, especiallysodium or calcium, or a solvate thereof, or a solvate of such a salt. Aparticular statin is atorvastatin, or a pharmaceutically acceptablesalt, solvate, solvate of such a salt or a prodrug thereof. A moreparticular statin is atorvastatin calcium salt. A particularly preferredstatin is, however, a compound with the chemical name(E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl(methylsulfonyl)-amino]-pyrimidin-5-yl](3R,5S)-3,5-dihydroxyhept-6-enoicacid, [also known as(E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[N-methyl-N-(methylsulfonyl)-amino]-pyrimidin-5-yl](3R,5S)-3,5-dihydroxyhept-6-enoicacid] or a pharmaceutically acceptable salt or solvate thereof, or asolvate of such a salt. The compound(E)-7-[4-(4-fluorophenyl)-6-isopropyl-2-[methyl-(methylsulfonyl)-amino]-pyrimidin-5-yl](3R,5S)-3,5-dihydroxyhept-6-enoicacid, and its calcium and sodium salts are disclosed in European PatentApplication, Publication No. EP-A-0521471, and in Bioorganic andMedicinal Chemistry, (1997), 5(2), 437-444. This latter statin is nowknown under its generic name rosuvastatin.

In the present application, the term “cholesterol-lowering agent” alsoincludes chemical modifications of the HMG-CoA reductase inhibitors,such as esters, prodrugs and metabolites, whether active or inactive.

The present invention also includes a compound of the present inventionin combination with a bile acid sequestering agent, for examplecolestipol or cholestyramine or cholestagel.

The present invention also includes a compound of the present inventionin combination with an inhibitor of the ileal bile acid transport system(IBAT inhibitor).

Suitable compounds possessing IBAT inhibitory activity have beendescribed, see for instance the compounds described in WO 93/16055, WO94/18183, WO 94/18184, WO 94/24087, WO 96/05188, WO 96/08484, WO96/16051, WO 97/33882, WO98/07749, WO 98/38182, WO 98/40375, WO98/56757, WO 99/32478, WO 99/35135, WO 99/64409, WO 99/64410, WO00/01687, WO 00/20392, WO 00/20393, WO 00/20410, WO 00/20437, WO01/34570, WO 00/35889, WO 00/47568, WO 00/61568, WO 01/68637, WO01/68096, WO 02/08211, WO 00/38725, WO 00/38726, WO 00/38727, WO00/38728, WO 00/38729, DE 19825804, JP 10072371, U.S. Pat. No.5,070,103, EP 251 315, EP 417 725, EP 489 423, EP 549 967, EP 573 848,EP 624 593, EP 624 594, EP 624 595, EP 869 121, EP 864 582, and EP 1 070703, and the contents of these patent applications, particularly thecompounds described in claim 1 and the named examples, are incorporatedherein by reference.

Particular classes of IBAT inhibitors suitable for use in the presentinvention are benzothiepines, and the compounds described in the claims,particularly claim 1, of WO 00/01687, WO 96/08484 and WO 97/33882 areincorporated herein by reference. Other suitable classes of IBATinhibitors are the 1,2-benzothiazepines, 1,4-benzothiazepines and1,5-benzothiazepines. A further suitable class of IBAT inhibitors is the1,2,5-benzothiadiazepines.

One particular suitable compound possessing IBAT inhibitory activity is(3R,5R)-3-butyl-3-ethyl-1,1-dioxido-5-phenyl-2,3,4,5-tetrahydro-1,4-benzothiazepin-8-ylβ-D-glucopyranosiduronic acid (EP 864 582). Other suitable IBATinhibitors include one of:

-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-1′-phenyl-1′-[N′-(carboxymethyl)    carbamoyl]methyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N′-(carboxymethyl)carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-1′-phenyl-1′-[N′-(2-sulphoethyl)carbamoyl]methyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3-butyl-3-ethyl-5-phenyl-7-methylthio-8-(N-{(R)-1′-phenyl-1′-[N′-(2-sulphoethyl)carbamoyl]methyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N′-(2-sulphoethyl)carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3-butyl-3-ethyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N′-(2-sulphoethyl)    carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3-butyl-3-ethyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N′-(2-carboxyethyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N′-(2-carboxyethyl)carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3-butyl-3-ethyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N′-(5-carboxypentyl)    carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N′-(2-carboxyethyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{α-[N′-(2-sulphoethyl)carbamoyl]-2-fluorobenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3-butyl-3-ethyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N′-(R)-(2-hydroxy-1-carboxyethyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N′-(R)-(2-hydroxy-1-carboxyethyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-{N-[(R)-α-(N′-{(R)-α-[N″-(R)-(2-hydroxy-1-carboxyethyl)carbamoyl]-2-hydroxyethyl}carbamoyl)benzyl]carbamoylmethoxy}-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3-butyl-3-ethyl-5-phenyl-7-methylthio-8-(N-{α-[N′-(carboxymethyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3-butyl-3-ethyl-5-phenyl-7-methylthio-8-(N-{α-[N′-((ethoxy)(methyl)phosphoryl-methyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3-butyl-3-ethyl-5-phenyl-7-methylthio-8-{N-[(R)-α-(N′-{2-[(hydroxy)(methyl)phosphoryl]ethyl}carbamoyl)benzyl]carbamoylmethoxy}-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N′-(2-methylthio-1-carboxyethyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-{N-[(R)-α-(N′-{2-[(methyl)(ethyl)    phosphoryl]ethyl}carbamoyl)-4-hydroxybenzyl]carbamoylmethoxy}-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-{N-[(R)-α-(N′-{2-[(methyl)(hydroxy)    phosphoryl]ethyl}carbamoyl)-4-hydroxybenzyl]carbamoylmethoxy}-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[(R)-N′-(2-methylsulphinyl-1-carboxyethyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methoxy-8-[N-{(R)-α-[N′-(2-sulphoethyl)carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy]-2,3,4,5-tetrahydro-1,5-benzothiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((R)-1-carboxy-2-methylthio-ethyl)carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((S)-1-carboxy-2-(R)-hydroxypropyl)carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((S)-1-carboxy-2-methylpropyl)carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((S)-1-carboxybutyl)    carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((S)-1-carboxypropyl)    carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((S)-1-carboxyethyl)    carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((S)-1-carboxy-2-(R)-hydroxypropyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-(2-sulphoethyl)carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((S)-1-carboxyethyl)carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((R)-1-carboxy-2-methylthioethyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-{(S)-1-[N-((S)-2-hydroxy-1-carboxyethyl)carbamoyl]propyl}carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((S)-1-carboxy-2-methylpropyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-((S)-1-carboxypropyl)    carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-[N-((R/S)-α-{N-[1-(R)-2-(S)-1-hydroxy-1-(3,4-dihydroxyphenyl)prop-2-yl]carbamoyl}-4-hydroxybenzyl)carbamoylmethoxy]-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;-   1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-(2-(S)-3-(R)-4-(R)-5-(R)-2,3,4,5,6-pentahydroxyhexyl)carbamoyl]-4-hydroxybenzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;    and-   1,1-Dioxo-3,3-dibutyl-5-phenyl-7-methylthio-8-(N-{(R)-α-[N-(2-(S)-3-(R)-4-(R)-5-(R)-2,3,4,5,6-pentahydroxyhexyl)carbamoyl]benzyl}carbamoylmethoxy)-2,3,4,5-tetrahydro-1,2,5-benzothiadiazepine;    or a pharmaceutically acceptable salt, solvate, solvate of such a    salt or a prodrug thereof.

According to an additional further aspect of the present invention thereis provided a combination treatment comprising the administration of aneffective amount of a compound of the formula I, or a pharmaceuticallyacceptable salt, solvate, solvate of such a salt or a prodrug thereof,optionally together with a pharmaceutically acceptable diluent orcarrier, with the simultaneous, sequential or separate administrationone or more of the following agents selected from:

-   a CETP (cholesteryl ester transfer protein) inhibitor, for example    those referenced and described in WO 00/38725 page 7 line 22-page    10, line 17 which are incorporated herein by reference;-   a cholesterol absorption antagonist for example azetidinones such as    SCH 58235 and those described in U.S. Pat. No. 5,767,115 which are    incorporated herein by reference;-   a MTP (microsomal transfer protein) inhibitor for example those    described in Science, 282, 751-54, 1998 which are incorporated    herein by reference;-   a nicotinic acid derivative, including slow release and combination    products, for example, nicotinic acid (niacin), acipimox and    niceritrol;-   a phytosterol compound for example stanols;-   probucol;-   an omega-3 fatty acid for example Omacor™;-   an anti-obesity compound for example orlistat (EP 129,748) and    sibutramine (GB 2,184,122 and U.S. Pat. No. 4,929,629);-   an antihypertensive compound for example an angiotensin converting    enzyme (ACE) inhibitor, an angiotensin II receptor antagonist, an    andrenergic blocker, an alpha andrenergic blocker, a beta    andrenergic blocker for example metoprolol, a mixed alpha/beta    andrenergic blocker, an andrenergic stimulant, calcium channel    blocker, an AT-1 blocker, a saluretic, a diuretic or a vasodilator;-   a CB1 antagonist or inverse agonist for example as described in    WO01/70700 and EP 65635;-   aspirin;-   a Melanin concentrating hormone (CH) antagonist;-   a PDK inhibitor; or    modulators of nuclear receptors for example LXR, FXR, RXR, and    RORalpha; or a pharmaceutically acceptable salt, solvate, solvate of    such a salt or a prodrug thereof, optionally together with a    pharmaceutically acceptable diluent or carrier to a warm-blooded    animal, such as man in need of such therapeutic treatment.

Particular ACE inhibitors or pharmaceutically acceptable salts,solvates, solvate of such salts or a prodrugs thereof, including activemetabolites, which can be used in combination with a compound of formulaI include but are not limited to, the following compounds: alacepril,alatriopril, altiopril calcium, ancovenin, benazepril, benazeprilhydrochloride, benazeprilat, benzoylcaptopril, captopril,captopril-cysteine, captopril-glutathione, ceranapril, ceranopril,ceronapril, cilazapril, cilazaprilat, delapril, delapril-diacid,enalapril, enalaprilat, enapril, epicaptopril, foroxymithine,fosfenopril, fosenopril, fosenopril sodium, fosinopril, fosinoprilsodium, fosinoprilat, fosinoprilic acid, glycopril, hemorphin-4,idrapril, imidapril, indolapril, indolaprilat, libenzapril, lisinopril,lyciumin A, lyciumin B, mixanpril, moexipril, moexiprilat, moveltipril,muracein A, muracein B, muracein C, pentopril, perindopril,perindoprilat, pivalopril, pivopril, quinapril, quinapril hydrochloride,quinaprilat, ramipril, ramiprilat, spirapril, spirapril hydrochloride,spiraprilat, spiropril, spiropril hydrochloride, temocapril, temocaprilhydrochloride, teprotide, trandolapril, trandolaprilat, utibapril,zabicipril, zabiciprilat, zofenopril and zofenoprilat. Preferred ACEinhibitors for use in the present invention are ramipril, ramiprilat,lisinopril, enalapril and enalaprilat. More preferred ACE inhibitors foruses in the present invention are ramipril and ramiprilat.

Preferred angiotensin II antagonists, pharmaceutically acceptable salts,solvates, solvate of such salts or a prodrugs thereof for use incombination with a compound of formula I include, but are not limitedto, compounds: candesartan, candesartan cilexetil, losartan, valsartan,irbesartan, tasosartan, telmisartan and eprosartan. Particularlypreferred angiotensin II antagonists or pharmaceutically acceptablederivatives thereof for use in the present invention are candesartan andcandesartan cilexetil.

Therefore in an additional feature of the invention, there is provided amethod for for the treatment of type 2 diabetes and its associatedcomplications in a warm-blooded animal, such as man, in need of suchtreatment which comprises administering to said animal an effectiveamount of a compound of formula I, or a pharmaceutically acceptablesalt, solvate, solvate of such a salt or a prodrug thereof insimultaneous, sequential or separate administration with an effectiveamount of one the other compounds described in this combination section,or a pharmaceutically acceptable salt, solvate, solvate of such a saltor a prodrug thereof.

Therefore in an additional feature of the invention, there is provided amethod of treating hyperlipidemic conditions in a warm-blooded animal,such as man, in need of such treatment which comprises administering tosaid animal an effective amount of a compound of formula I, or apharmaceutically acceptable salt, solvate, solvate of such a salt or aprodrug thereof in simultaneous, sequential or separate administrationwith an effective amount of one the other compounds described in thiscombination section or a pharmaceutically acceptable salt, solvate,solvate of such a salt or a prodrug thereof.

According to a further aspect of the invention there is provided apharmaceutical composition which comprises a compound of formula I, or apharmaceutically acceptable salt, solvate, solvate of such a salt or aprodrug thereof, and one of the other compounds described in thiscombination section or a pharmaceutically acceptable salt, solvate,solvate of such a salt or a prodrug thereof, in association with apharmaceutically acceptable diluent or carrier.

According to a further aspect of the present invention there is provideda kit comprising a compound of formula I, or a pharmaceuticallyacceptable salt, solvate, solvate of such a salt or a prodrug thereof,and one of the other compounds described in this combination section ora pharmaceutically acceptable salt, solvate, solvate of such a salt or aprodrug thereof.

According to a further aspect of the present invention there is provideda kit comprising:

-   a) a compound of formula I, or a pharmaceutically acceptable salt,    solvate, solvate of such a salt or a prodrug thereof, in a first    unit dosage form;    -   b) one of the other compounds described in this combination        section or a pharmaceutically acceptable salt, solvate, solvate        of such a salt or a prodrug thereof; in a second unit dosage        form; and-   c) container means for containing said first and second dosage    forms.

According to a further aspect of the present invention there is provideda kit comprising:

-   a) a compound of formula I, or a pharmaceutically acceptable salt,    solvate, solvate of such a salt or a prodrug thereof, together with    a pharmaceutically acceptable diluent or carrier, in a first unit    dosage form;-   b) one of the other compounds described in this combination section    or a pharmaceutically acceptable salt, solvate, solvate of such a    salt or a prodrug thereof, in a second unit dosage form; and-   c) container means for containing said first and second dosage    forms.

According to another feature of the invention there is provided the useof a compound of the formula I, or a pharmaceutically acceptable salt,solvate, solvate of such a salt or a prodrug thereof, and one of theother compounds described in this combination section, or apharmaceutically acceptable salt, solvate, solvate of such a salt or aprodrug thereof, in the manufacture of a medicament for use in the thetreatment of metabolic syndrome or type 2 diabetes and its associatedcomplications in a warm-blooded animal, such as man.

According to another feature of the invention there is provided the useof a compound of the formula I, or a pharmaceutically acceptable salt,solvate, solvate of such a salt or a prodrug thereof, and one of theother compounds described in this combination section, or apharmaceutically acceptable salt, solvate, solvate of such a salt or aprodrug thereof, in the manufacture of a medicament for use in thetreatment of hyperlipidaemic conditions in a warm-blooded animal, suchas man.

According to a further aspect of the present invention there is provideda combination treatment comprising the administration of an effectiveamount of a compound of the formula I, or a pharmaceutically acceptablesalt, solvate, solvate of such a salt or a prodrug thereof, optionallytogether with a pharmaceutically acceptable diluent or carrier, with thesimultaneous, sequential or separate administration of an effectiveamount of one of the other compounds described in this combinationsection, or a pharmaceutically acceptable salt, solvate, solvate of sucha salt or a prodrug thereof, optionally together with a pharmaceuticallyacceptable diluent or carrier to a warm-blooded animal, such as man inneed of such therapeutic treatment.

EXAMPLES

¹H NMR and ¹³C NMR measurements were performed on a Varian Mercury 300or Varian UNITY plus 400, 500 or 600 spectrometers, operating at ¹Hfrequencies of 300, 400, 500 and 600 MHz, respectively, and at ¹³Cfrequencies of 75, 100, 125 and 150 MD, respectively. Measurements weremade on the delta scale (δ).

Unless otherwise stated, chemical shifts are given in ppm with thesolvent as internal standard. Abbreviations IRS insulin resistancesyndrome TLC thin layer chromatography HOBT1-hydroxybenzotriazole-hydrate DIBAH diisobutylaluminium hydride DMSOdimethyl sulfoxide EtOAc ethyl acetate DMF N,N-dimethylformamide THFtetrahydrofuran PEG polyethylene glycol HPLC high performance liquidchromatography MeCN acetonitrile TFA trifluoroacetic acid Pd/C palladiumon charcoal HATU O-(7-azabenzotriazolyl-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate DCM dichloromethane TBTUO-(benzotriazol-1-yl)-N,N,N′,N′- tetramethyluronium tetrafluoborateDIPEA N,N-diisopropylethylamine DMAP 4-dimethylaminopyridine TrisamineTris(hydroxymethyl)aminomethane ISOLUTE ® FLASH Si is a silica columnsuitable for chromatography Borohydride on polymer support isBorohydride on Amberlite IRA-400 available from Aldrich EDC1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride NH₄OAcammonium acetate t triplet s singlet d doublet q quartet qvint quintet mmultiplet br broad bs broad singlet dm doublet of multiplet bt broadtriplet dd doublet of doublet

Example 1 a) N-benzyl-N-hexyl-3-(4-hydroxyphenyl)propanamide

N-hexylbenzylamine (0.6 g, 3.136 mmol) and 3-(4-hydroxyphenyl)propionicacid (0.52 g 3.136 mmol) were mixed in DMF (10 ml) and the mixture wascooled. HOBT (0.424 g, 3.136 mmol) and the TBTU (1 g, 3.136 mmol)reagent were added followed by DIPEA (1.216 g, 9.409 mmol). The mixturewas stirred at room temperature overnight and then evaporated. Theresulting mixture was partitioned between ethyl acetate and sodiumhydrogencarbonate aqueous solution (sat.). The aqueous portion wasextracted with ethyl acetate and the combined organic extracts driedwith magnesium sulphate and then evaporated. Chromatography of theresidue on a column (ISOLUTE® SI, 5 g/25 ml) using ethyl acetate/heptane(10:90, then 25:75) as eluant gave 760 mg the desired product, yield71%.

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.84 (t, 3H), 1.16-1.27 (m, 6H),1.41-1.51 (m, 2H), 2.55, 2.63 (t, t, 2H), 2.88, 2.94 (t, t, 2H), 3.09,3.31 (t, t, 2H), 4.40, 4.57 (s, s, 2H), 6.69, 6.73 (d, d, 2H), 6.98 (d,2H), 7.05, 7.07 (d, d, 2H), 7.14 (d, 1H) and 7.21-7.31 (m, 5H)

b) Methyl2-[(4-{3-[benzyl(hexyl)amino]-3-oxopropyl}phenoxy)methyl]benzoate

N-benzyl-N-hexyl-3-(4-hydroxyphenyl)propanamide (183 mg, 0.54 mmol),methyl 2-(bromomethyl)benzoate (136 mg, 0.59 mmol) and potassiumcarbonate (112 mg, 0.81 mmol) were mixed in acetonitrile. The mixturewas stirred at 66° C. overnight. The solvent was evaporated underreduced pressure and the residue was dissolved in ethyl acetate. Theorganic phase was washed (water x2, brine x1), dried (Na₂SO₄) andevaporated. Further purification by preparative HPLC (using a gradientof CH₃CN/10% CH₃CN-waterphase containing 0.1M NH₄OAc as eluant) gave 91mg (yield 34%) of the desired product. ¹HNMR (rotamers, 400 MHz, CDCl₃):δ 0.84-0.88 (m, 3H), 1.19-1.29 (m, 6H), 1.42-1.53 (m, 2H), 2.57, 2.65(t, t, 2H), 2.92, 2.99 (t, 2H), 3.10, 3.34 (t, t, 2H), 3.89, 3.90 (s, s,3H), 4.42, 4.60 (s, s, 2H), 5.47, 5.48 (s, s, 2H), 6.86-6.93 (m, 2H),7.07 (t, 2H), 7.14-7.19 (m, 2H), 7.21-7.38 (m, 4H), 7.52-7.56 (m, 1H),7.75 (t, 1H), 8.00-8.03 (m, 1H).

c) 2-[(4-{3-[Benzyl(hexyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic acid

Methyl 2-[(4-{3-[benzyl(hexyl)amino]-3-oxopropyl}phenoxy)methyl]benzoate(61 mg, 0.13 mmol) and lithium hydroxide (7 mg, 0.29 mmol) weredissolved in 3 ml of a 1:1 mixture of THF and water in a microwave vial.The resulting reaction mixture was irradiated in a microwave oven at120° C. for 40 min.

Water was added and the THF was evaporated under reduced pressure. Theresidue was acidified with 1M hydrochloric acid and extracted with ethylacetate (x3). The organic phases were combined, washed (water, brine),dried (Na₂SO₄) and evaporated. The crude product was further purified bypreparative HPLC (using a gradient of CH3CN/10% CH₃CN-waterphasecontaining 0.1M NH₄OAc as eluant). 38 mg (yield 64%) of pure product wasobtained after freeze-drying.

¹HNMR (rotamers, 400 MHz, CDCl₃): δ 0.86 (t, 3H), 1.19-1.28 (m, 6H),1.43-1.55 (m, 2H), 2.60, 2.69 (t, t, 2H), 2.92, 2.99 (t, t, 2H), 3.11,3.36 (t, t, 2H), 4.43, 4.61 (s, s, 2H), 5.52, 5.53 (s, s, 2H), 6.87-6.93(m, 2H), 7.05-7.09 (m, 2H), 7.14-7.33 (m, 5H), 7.36-7.40 (m, 1H),7.55-7.59 (m, 1H), 7.77 (t, 1H) and 8.12-8.15 (m, 1H).

¹³CNMR (rotamers, 100 MHz, CDCl₃): δ 14.17, 14.23, 22.73, 22.79, 26.75,26.88, 27.68, 27.72, 31.11, 31.26, 31.61, 31.82, 35.39, 35.68, 46.84,47.48, 48.72, 51.35, 68.42, 115.16, 115.22, 126.39, 127.15, 127.42,127.49, 127.72, 128.23, 128.70, 129.10, 129.72, 131.81, 133.45, 133.81,133.84, 137.10, 137.82, 140.88, 157.33, 157.40, 171.20, 173.10, 173.39.

Example 2 a) N-(2,4-difluorobenzyl)-N-heptylamine

Heptylamine (345.6 mg, 3 mmol) was added into 2,4-difluorobenzaldehyde(440.5 mg, 3.1 mmol) in MeOH (3 ml) and trimethyl orthoformate (2 ml),followed by acetic acid (0.05 ml). The mixture was in microwave oven(Smith Synthesizer) at 150° C. for 10 minutes. DCM (3 ml) was then addedand followed borohydride on polymer support (1.2 g, 3 mmol). The mixturewas shaken overnight and more of borohydride on polymer support (1.2 g)was added. The mixture was shaken over weekend and then filtered andevaporated. The residue was put on a column ((ISOLUTE®PRS, 10 g) andeluted with MeCN, MeOH and then MeOH(NH₃ sat.). 536 mg of oil productwas obtained, yield 72%. ¹H NMR (400 MHz, CDCl₃): δ 0.87 (t, 3H),1.23-1.32 (m, 8H), 1.45-1.52 (m, 2H), 2.59 (t, 2H), 3.78 (s, 2H),6.75-6.85 (m, 2H) and 7.27-7.33 (m, 1H)

b) (4-{[2-(Methoxycarbonyl)benzyl]thio}phenyl)acetic acid

4-Mercaptophenylacetic acid (995 mg, 5.915 mmol) in THF (15 ml) wascooled in an ice-bath and sodium hydride (55-65%, 520 mg, ˜13 mmol) wasadded. The mixture was stirred for 30 minutes and then2-bromomethyl-benzoic acid methyl ester (1.49 g, 6.507 mmol) in THF (5ml) was added. The resulting mixture was stirred overnight and thetemperature was allowed going up to room temperature. Water was droppedin and the mixture was stirred for ca. 20 minutes. It was thenevaporated to remove THF. The residue was acidified with 1% hydrochloricacid, pH˜3, and then extracted with ethyl acetate. The organic extractswere combined, dried with magnesium sulphate and evaporated.Chromatography of the residue on a column (ISOLUTE®SI, 20 g/70 ml) usingDCM, then MeOH/DCM (1:99) as eluant gave 224 mg desired product, yield65%. ¹H NMR (500 MHz, CDCl₃): δ 3.62 (s, 2H), 3.90 (s, 3H), 4.52 (s,2H), 7.17 (d, 2H), 7.23-7.40 (m, 5H) and 7.94 (d, 1H)).

c) Methyl2-{[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]-methyl}benzoate

(4-{[2-(Methoxycarbonyl)benzyl]thio}phenyl)acetic acid (581 mg, 1.836mmol) and N-(2,4-difluorobenzyl)-N-heptylamine (465.3 mg, 1.968 mmol)were combined in DMF and the mixture was cooled in an ice-bath. HOBT(260.6 mg, 1.928 mmol) and TBTU (619 mg, 1.928 mmol) were added,followed by DIPEA (747.7 mg 5.785 mmol). The mixture was stirred at roomtemperature overnight and then evaporated. The resulting mixture waspartitioned between ethyl acetate and sodium hydrogencarbonate aqueoussolution (sat.). The aqueous portion was extracted with ethyl acetateand the combined organic extracts was dried with magnesium sulphate andthen evaporated. Chromatography of the residue on a column (ISOLUTE®SI,20 g/70 ml) using ethyl acetate/heptane (5:95, then 10:90) as eluantgave 767 mg the desired product, yield 77%.

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.88-0.93 (m, 3H), 1.23-1.34 (m,8H), 1.48-1.57 (m, 2H), 3.19-3.24, 3.30-3.37 (m, m, 2H), 3.67-3.74 (m,2H), 3.92 (s, 3H), 4.50, 4.63 (s, s, 2H), 4.53 (s, 2H), 6.78-6.89 (m,2H), 7.00-7.40 (m, 8H) and 7.95 (d, 2H).

d)2-{[(4-{2-[(2,4-Difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoicacid

Methyl2-{[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]methyl}-benzoate(31 mg, 0.057 mmol) was dissolved in THF (1 ml) and cooled in anice-bath. Lithium hydroxide (2 mg, 0.075 mmol) in water (1 ml) wasadded. After the addition, the cooling bath was removed and the mixturewas stirred overnight. LC-MS showed there was very little product. Morelithium hydroxide (3 mg) was added and the mixture was stirred for 6days more and HPLC showed about 30% product. More (3 mg) of lithiumhydroxide was added and the mixture was stirred for 13 days more. It wasevaporated in vacuum to remove THF. The residue was acidified with 10%hydrochloric acid, pH˜3, and extracted with ethyl acetate (x2). Theorganic phases were combined, dried (magnesium sulphate) and evaporated.Chromatography of the residue on a column (ISOLUTE® SI, 500 g/3 ml)using DCM, MeOH/DCM (0.5:99.5) as eluant gave 17 mg desired product,yield 56%.

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.89-0.93 (m, 3H), 1.25-1.34 (m,8H), 1.52-1.60 (m, 2H), 3.27, 3.37 (t, t, 2H), 3.71, 3.76 (s, s, 2H),4.55, 4.65 (s, s, 2H), 6.80-6.92 (m, 2H), 7.07-7.17 (m, 2H), 7.28-7.37(m, 5H), 7.42-7.47 (m, 1H) and 7.95-7.98 (m, 1H).

¹³C NMR (rotamers, 125 MHz, CDCl₃): δ 14.01, 22.51, 26.78, 26.85, 27.28,28.56, 28.85, 28.93, 31.65, 31.70, 38.22, 39.94, 40.25, 41.57, 41.60,45.16, 46.48, 47.94, 103.53 (t), 104.26(t), 111.50(br), 111.71(br),119.65(d), 120.37(d), 127.05, 128.95 (br), 129.61, 129.68, 131.15,131.66(d), 131.76(d), 132.41, 132.80, 132.88, 133.85, 134.03, 140.43,160.58 (dd), 160.91(dd), 162.21(dd), 162.53(dd), 170.46, 171.44 and171.54.

Example 3 a) (4-{[2-(Methoxycarbonyl)benzyl]oxy}phenyl)acetic acid

4-Hydroxyphenylacetic acid (760 mg, 4.995 mmol) was dissolved in ethanol(99.5%, 20 ml). Potassium hydroxide (560.5 mg, 9.99 mmol) was added. Themixture was stirred at room temperature for 30 minutes.2-Bromomethylbenzoic acid methyl ester (1144.2 mg, 4.995 mmol) was thendropped in. The resulting mixture was heated to reflux for 2 hours andthen evaporated in vacuum to dry. Water and ethyl acetate were addedinto the residue and the phases were separated. The water phase wasacidified with 10% hydrochloric acid, pH˜5, and then extracted withethyl acetate. The organic phase was dried with magnesium sulphate andevaporated in vacuum to dry. Chromatography of the residue on a column(ISOLUTE® SI, 5 g/6 ml) using DCM, MeOH/DCM (1:99) as eluant gave thedesired product (262 mg), yield 17.5%.

¹H NMR (500 MHz, CDCl₃): δ 3.61 (s, 2H), 3.91 (s, 3H), 5.50 (s, 2H),6.97 (d, 2H), 7.22 (d, 2H), 7.39 (t, 1H), 7.57 (t, 1H), 7.76 (d, 1H) and8.04 (d, 1H).

b) Methyl2-[(4-{2-[benzyl(hexyl)amino]-2-oxoethyl}phenoxy)methyl]benzoate

(4-{[2-(Methoxycarbonyl)benzyl]oxy}phenyl)acetic acid (50 mg, 0.166mmol) was dissolved in DCM (2 ml), N-hexylbenzylamine (38.2 mg, 0.2mmol) was added, then EDC (38.3 mg, 0.2 mmol) was added and then DMAP(24.4 mg, 0.2 mmol) was added. The mixture was stirred at roomtemperature overnight. 1% HCl (1 ml) and water (1 ml) were added intothe mixture. The two phases were separated by using a Whatman FilterTube. The obtained organic solution was evaporated in vacuum and the oilproduct (71 mg) was left. It was then used directly in next step.

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.86-0.91 (m, 3H), 1.22-1.32 (m,6H), 1.47-1.58 (m, 2H), 3.21, 3.39 (t, t, 2H), 3.65, 3.75 (s, s, 2H),3.93 (s, 3H), 4.53, 4.64 (s, s, 2H), 5.51, 5.52 (s, s, 2H), 6.96, 6.99(d, d, 2H), 7.16 (d, 2H), 7.23-7.42 (m, 6H), 7.59 (t, 1H), 7.78 (d, 1H)and 8.06 (d, 1H).

c) 2-[(4-{2-[Benzyl(hexyl)amino]-2-oxoethyl}phenoxy)methyl]benzoic acid

Methyl 2-[(4-{2-[benzyl(hexyl)amino]-2-oxoethyl}phenoxy)methyl]benzoate(70 mg, 0.148 mmol) in THF (2 ml) was cooled in an ice-bath. Lithiumhydroxide (7.08 mg, 0.296 mmol) in water (1 ml) was dropped in. Thecooling-bath was then removed and the mixture was stirred overnight.HPLC showed that the reaction was not complete. More lithium hydroxide(0.2M, 0.5 ml) was added. The reaction mixture was stirred for 4 daysmore. It was then evaporated in vacuum to remove THF. The residue wasacidified with 1% hydrochloric acid, pH=3-4, and extracted with ethylacetate. The organic phase was dried (magnesium sulphate) andevaporated. Chromatography of the residue on a column (ISOLUTE® SI, 2g/6 ml) using DCM, MeOH/DCM (1:99, and then 2:98) as eluant gave 24 mgthe desired product, yield 35%.

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.86-0.90 (m, 3H), 1.21-1.30 (m,6H), 1.46-1.58 (m, 2H), 3.21, 3.39 (t, t, 2H), 3.68, 3.77 (s, s, 2H),4.53, 4.65 (s, s, 2H), 5.53, 5.54 (s, s, 2H), 6.95, 6.98 (d, d, 2H),7.14-7.17 (m, 2H), 7.22-7.33 (m, 4H), 7.35-7.43 (m, 2H), 7.60 (t, 1H),7.80 (d, 1H) and 8.16 (d, 1H).

¹³C NMR (rotamers, 125 MHz, CDCl₃): δ 13.94, 13.97, 22.49, 22.53, 26.47,26.57, 27.29, 28.36, 31.40, 31.50, 39.82, 40.12, 46.50, 47.43, 48.28,51.31, 68.21, 115.15, 126.25, 126.85, 127.20, 127.24, 127.36, 127.48,127.54, 127.97, 128.49, 128.87, 129.76, 129.89, 131.52, 133.18, 136.80,137.57, 140.40, 157.59, 170.60, 171.76 and 172.03.

Example 4 a) Methyl2-[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenoxy)methyl]-benzoate

N-(2,4-Difluorobenzyl)-N-heptylamine (106 mg, 0.44 mmol) was added into(4-{[2-(methoxycarbonyl)benzyl]oxy}phenyl)acetic acid (120 mg, 0.4 mmol)in DCM (10 ml) and followed by EDC (84.3 mg, 0.44 mmol) and then DMAP(54 mg, 0.44 mmol). The mixture was stirred at room temperatureovernight, and then washed with 1% hydrochloric acid, water and brineand dried with magnesium sulphate and evaporated. Chromatography of theresidue on a column (ISOLUTE®SI, 5 g/15 ml) using DCM and MeOH/DCM(0.5:99.5) as eluant gave 155 mg desired product, yield 74%.

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.88-0.92 (m, 3H), 1.23-1.33 (m,8H), 1.49-1.57 (m, 2H), 3.24, 3.34 (t, t, 2H), 3.66, 3.72 (s, s, 2H),3.92 (s, 3H), 4.53, 4.62 (s, s, 2H), 5.50, 5.51 (s, s, 2H), 6.77-6.89(m, 2H), 6.95, 6.98 (d, d, 2H), 6.99-7.04, 7.29-7.33 (m, m, 1H), 7.17,7.20 (d, d, 2H), 7.39 (t, 1H), 7.57 (t, 1H), 7.75-7.79 (m, 1H) and 8.05(d, 1H).

b)2-[(4-{2-[(2,4-Difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenoxy)methyl]benzoicacid

Lithium hydroxide (13.3 mg, 0.554 mmol) in water (1.5 ml) was added into70335 methyl2-[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenoxy)methyl]benzoate(145 mg, 0.277 mmol) dissolved in THF (3 ml). The mixture was then inmicrowave oven (Smith Synthesizer) at 150° C. for 7 minutes and thenevaporated to remove THF. The residue was acidified with 1% hydrochloricacid, pH˜4 and then extracted with ethyl acetate (x2). The organicportions were combined, washed with brine, dried with magnesium sulphateand then evaporated. Chromatography of the residue on a column (ISOLUTE®SI, 2 g/6 ml) using DCM then MeOH/DCM (0.5:99.5, then 1:99) as eluantgave 94 mg desired product, yield 67%.

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.86-0.91 (m, 3H), 1.22-1.32 (m,8H), 1.48-1.56 (m, 2H), 3.23, 3.34 (t, t, 2H), 3.68, 3.73 (s, s, 2H),4.52, 4.63 (s, s, 2H), 5.53 (s, br, 2H), 6.77-6.87 (m, 2H), 6.93-6.97(m, 2H), 6.99-7.04, 7.27-7.33 (m, m, 1H), 7.16-7.20 (m, 2H), 7.41 (t,1H), 7.58-7.62 (m, 1H), 7.78 (d, 1H) and 8.15 (d, 1H).

¹³C NMR (rotamers, 125 MHz, CDCl₃): δ 14.00, 22.51, 26.75, 26.84, 27.26,28.54, 28.86, 28.94, 31.64, 31.70, 39.75, 40.08, 41.39, 45.05, 46.30,47.98, 68.21, 103.67 (t), 104.12 (t), 111.52 (d), 115.17, 119.80 (d),120.52 (d), 126.97, 127.04, 127.21, 128.81 (br), 129.78, 129.85, 131.52,133.16, 140.33, 157.65, 160.48 (dd), 160.85 (dd), 162.13 (dd), 162.46(dd), 171.01, 171.93 and 171.99.

Example 5 a)N-(2,4-difluorobenzyl)-N-heptyl-3-(4-hydroxyphenyl)propanamide

3-(4-Hydroxyphenyl)propionic acid (108 mg, 0.650 mmol) was dissolved inDMF. N-(2,4-difluorobenzyl)-N-heptylamine (164.7 mg, 0.682 mmol) wasadded. The mixture was cooled in an ice-bath. TBTU (219 mg, 0.682 mmol)was added and followed by DIPEA (0.238 ml, 1.365 mmol). The mixture wasstirred overnight and the temperature was allowed up to roomtemperature. Ethyl acetate and sodium hydrogencarbonate aqueous solution(sat.) were added and then the two phases were separated. The waterphase was extracted with ethyl acetate. The organic phases were combinedand dried with magnesium sulphate and evaporated. Chromatography of theresidue on a column (ISOLUTE® SI, 5 g/15 ml) using DCM and then MeOH/DCM(1:99) as eluant gave 223 mg desired product, yield 88%.

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.86-0.90 (m, 3H), 1.21-1.31 (m,8H), 1.47-1.53 (m, 2H), 2.60, 2.67 (t, t, 2H), 2.85-2.96 (m, 2H), 3.15,3.32 (t, t, 21), 4.41, 4.60 (s, s, 2H), 6.75-6.85 (m, 4H), 6.90-6.96,7.12-7.18 (m, m, 1H) and 7.00, 7.04 (d, d, 2H).

b) Methyl2-[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenoxy)methyl]-benzoate

N-(2,4-difluorobenzyl)-N-heptyl-3-(4-hydroxyphenyl)propanamide (195 mg,0.501 mmol), 2-bromomethylbenzoic acid methyl ester (120.4 mg, 0.526mmol) and anhydrous potassium carbonate (103 mg, 0.751 mmol) were mixedin acetonitrile (15 ml). The mixture was heated to reflux overnight andthen evaporated to dryness. Water and ethyl acetate were added and thetwo phases were separated. The organic phase was dried (magnesiumsulphate) and evaporated. Chromatography of the residue on a column(ISOLUTE® SI, 2 g/6 ml) using heptane/DCM (50:50), then DCM and thenMeOH/DCM (0.5:99.5) as eluant gave 187 mg desired product, yield 69.5%.

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.87-0.91 (m, 3H), 1.21-1.31 (m,8H), 1.44-1.56 (m, 2H), 2.56-2.69 (m, 2H), 2.91-3.01 (m, 2H), 3.14, 3.32(t, t, 2H), 3.92 (s, 3H), 4.43, 4.59 (s, s, 2H), 5.49 (s, 2H), 6.75-6.97(m, 4H), 7.08-7.28 (m, 3H), 7.38 (t, 1H), 7.56 (t, 1H), 7.76 (d, 1H) and8.04 (d, 1H),

c)2-[(4-{3-[(2,4-Difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid

Lithium hydroxide (13.3 mg, 0.554 mmol) in water (1 ml) was added intomethyl2-[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenoxy)methyl]benzoate(149 mg, 0.277 mmol) dissolved in THF (2 ml). The mixture was thenplaced in microwave oven (Smith Synthesizer) at 150° C. for 7 minutesand then evaporated to remove THF. The residue was acidified with 1%hydrochloric acid, pH˜4, and extracted with ethyl acetate (x2). Theorganic extracts were combined and washed with brine and dried withmagnesium sulphate and then evaporated. Chromatography of the residue ona column (ISOLUTE® SI, 2 g/6 ml) using DCM, then MeOH/DCM (1:99) aseluant gave 121 mg desired product, yield 83%:

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.89-0.93 (m, 3H), 1.24-1.33 (m,8H), 1.49-1.58 (m, 2H), 2.64-2.74 (m, 2H), 2.97-3.03 (m, 2H), 3.17, 3.37(t, t, 2H), 4.46, 4.65 (s, s, 2H), 5.58, 5.59 (s, s, 2H), 6.78-6.87 (m,2H), 6.94-6.97 (m, 2H), 6.99-7.04, 7.27-7.31 (m, m, 1H), 7.14, 7.17 (d,d, 2H), 7.42-7.45 (m, 1H), 7.61-7.64 (m, 1H), 7.82-7.85 (m, 1H) and8.19-8.22 (d, 1H),

¹³C NMR (rotamers, 125 MHz, CDCl₃): δ 13.96, 22.46, 22.49, 26.67, 26.85,27.38, 28.57, 28.80, 28.94, 30.69, 30.84, 31.58, 31.67, 35.01, 35.26,41.63, 44.76, 44.78, 46.43, 47.78, 68.15, 103.60(t), 104.07(t),111.41(dd), 111.49 (dd), 114.91, 119.74 (d), 120.45 (d), 126.86, 127.12,128.53 (br), 129.40, 131.49, 131.58, 133.15, 133.32, 140.54, 157.12,160.33 (dd), 160.81 (dd), 162.07 (dd), 162.33 (dd), 171.07, 173.04 and173.11.

Example 6

a) 3-(4-Mercaptophenyl)propanoic acid (2.0 g, 10.97 mmol) was dissolvedin dry THF (60 ml) and cooled to 0° C. Sodium hydride (0.64 g, 24.1mmol) was added. The mixture was stirred for 30 minutes, methyl2-(bromomethyl)benzoate (2.77 g, 12.07 mmol) dissolved in dry THF (10ml) was added dropwise. The solution was allowed to warm up to roomtemperature and was stirred overnight. Dropwise addition of water (10ml) deactivated the remaining sodium hydride. The solvent was removed byevaporation, and the residue was acidified to pH 3 (HCl 1%). The waterphase was washed with EtOAc (3×10 ml). The organic phases was combined,dried (MgSO₄) and evaporated. The crude was purified by preparative HPLC(started with acetonitrile/buffer 60/40 and then the acetonitrileconcentration was increased to 100%, the buffer was a mixture ofacetonitrile/water 10/90 and ammonium acetate (0.1 M, columnKR-100-7-C8, 50 mm×250 mm, flow 40 ml/min). The product containingfractions was pooled and the acetonitrile was removed by evaporation.EtOAc (10 ml) was added and the organic phase was washed with twopotions of brine and dried (MgSO₄). Removing the solvent by evaporationgave 2.26 gram of 3-(4-{[2-(methoxycarbonyl)benzyl]thio}phenyl)propanoicacid (yield 62.3%).

¹H NMR (500 MHz, CDCl₃): δ 2.66 (t, 2H), 2.92 (t, 2H), 3.90 (s, 3H),4.51 (s, 2H), 7.10 (d, 2H), 7.19 (d, 1H), 7.25 (d, 2H), 7.29 (t, 1H),7.36 (t, 1H) and 7.94 (d, 1H).

b) Methyl2-{[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenyl)thio]-methyl}benzoate

N-(2,4-difluorobenzyl)-N-heptylamine (0.64 g, 2.65 mmol) was dissolvedin DMF (10 ml), 3-(4-{[2-(methoxycarbonyl)benzyl]thio}phenyl)propanoicacid (0.80 g, 2.41 mmol) was added and the mixture was cooled to 0° C.N-[(1H-1,2,3-benzotriazol-1-yloxy)-(dimethylamino)methylene]-N-methylmethanaminiumtetrafluoroborate (0.85 g, 2.65 mmol) and diisopropylethylamine (0.65 g,5.05 mmol) was added. The mixture was allowed to warm to roomtemperature and stirred overnight. EtOAc (15 ml) was added and theorganic phase was washed with two portions of sodium hydrogencarbonate(aq, 10 ml). EtOAc was removed by evaporation and the crude was purifiedby preparative HPLC (started with acetonitrile/buffer 60/40 and then theacetonitrile concentration was increased to 100%, the buffer was amixture of acetonitrile/water 10/90 and ammonium acetate (0.1 M, columnKR-100-7-C8, 50 mm×250 mm, flow 40 ml/min). The product containingfractions were pooled and the acetonitrile was removed by evaporation.EtOAc (10 ml) was added and the organic phase was washed with twoportions of brine and dried (MgSO₄). The solvent was removed byevaporation and gave 1.10 gram of methyl2-{[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenyl)thio]methyl}benzoate(yield 82.2%).

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.89 (t, 3H), 1.22-1.32 (m, 8H),1.47-1.55 (m, 2H), 2.55, 2.66 (t, t, 2H), 2.95-3.01 (m, 2H), 3.16, 3.33(t, t, 2H), 3.89, 3.90 (s, s, 3H), 4.44, 4.60 (s, s, 2H), 4.50, 4.51 (s,s, 2H), 6.76-6.85 (m, 2H), 6.92-6.96, 7.20-7.25 (m, m, 4H), 7.07, 7.12(d, d, 2H), 7.27-7.31 (m, 1H), 7.32-7.34 (m, 1H) and 7.91-7.92 (m, 1H).

c)2-{[(4-{3-[(2,4-Difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenyl)thio]-methyl}benzoicacid

Methyl2-{[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenyl)thio]-methyl}benzoate(1.05 g, 1.89 mmol) was dissolved in EtOH (95%, 5 ml), potassiumhydroxide (0.21 g, 3.77 mmol) was added. The reaction was performed in asingle node microwave oven (7 min, 150° C.). Workup was by addition ofEtOAc (5 ml) and washing with HCl (2×5 ml, 1M). The organic layer wasdried (MgSO₄) and the solvent was removed by evaporation to give 0.96gram of2-{[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenyl)thio]methyl}benzoicacid (yield 94.3%).

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.88-0.92 (m, 3H), 1.24-1.33 (m,8H), 1.50-1.57 (m, 2H), 2.64, 2.69 (t, t, 2H), 2.95-3.00 (m, 2H), 3.20,3.34 (t, t, 2H), 4.48, 4.62 (s, s, 2H), 4.55, 4.56 (s, s, 2H), 6.79-6.87(m, 2H), 6.98-7.03, 7.27-7.30 (m, m, 2H), 7.06-7.10 (m, 2H), 7.22-7.24(m, 2H), 7.31-7.36 (m, 1H), 7.41-7.46 (m, 1H) and 8.03 (d, 1H).

¹³C NMR (rotamers, 125 MHz, CDCl₃): δ 14.01, 22.51, 22.54, 26.74, 26.90,27.42, 28.63, 28.86, 28.98, 30.92, 31.18, 31.64, 31.72, 34.65, 38.23,38.28, 41.58, 44.82, 46.44, 47.77, 103.46 (t), 104.14(t), 111.52(dd),111.58(dd), 119.74(dd), 120.51(dd), 127.09, 128.55, 128.63, 129.03,131.16, 131.49, 131.55, 131.78 (dd), 132.30, 132.50, 132.96, 140.04,140.11, 140.59, 140.68, 160.47(dd), 160.88(dd), 162.16(dd), 162.44(dd),170.88(br), 172.56 and 172.59.

Example 7

a) N-(2,3-dimethoxybenzyl)butan-1-amine (0.59 g, 2.65 mmol) wasdissolved in DMF (10 ml), 3-(4-hydroxyphenyl)propanoic acid (0.4 g, 2.41mmol), was added and the mixture was cooled to 0° C.N-[(1H-1,2,3-benzotriazol-1-yloxy)(dimethylamino)methylene]-N-methylmethanaminiumtetrafluoroborate (0.85 g, 2.65 mmol) and diisopropylethylamine (0.65 g,5.05 mmol) was added. The mixture was allowed to warm to roomtemperature and stirred over night. EtOAc (15 ml) was added and theorganic phase was washed with two portions of sodium hydrogencarbonate(aq, 10 ml). EtOAc was removed by evaporation and the crude was purifiedby preparative HPLC (started with acetonitrile/buffer 60/40 and then theacetonitrile concentration was increased to 100%, the buffer was amixture of acetonitrile/water 10/90 and ammonium acetate (0.1 M, columnKR-100-7-C8, 50 mm×250 mm, flow 40 ml/min). The product containingfractions was pooled and the acetonitrile was removed by evaporation.EtOAc (10 ml) was added and the organic phase was washed with twopotions of brine and dried (MgSO₄). The solvent was removed byevaporation and gave 1.08 gram ofN-butyl-N-(2,3-dimethoxybenzyl)-3-(4-hydroxyphenyl)propanamide (yield82.3%).

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.85-0.89 (m, 3H), 1.20-1.30 (m,2H), 1.44-1.53 (m, 2H), 2.60, 2.65 (t, t, 2H), 2.88, 2.94 (t, t, 2H),3.13, 3.33 (t, t, 2H), 3.78, 3.80 3.83, 3.85 (s, s, s, s, 6H), 4.43,4.68 (s, s, 2H), 6.57, 6.67 (d, d, 1H), 6.74-6.86 (m, 3H) and 6.95-7.05(m, 3H).

b) N-butyl-N-(2,3-dimethoxybenzyl)-3-(4-hydroxyphenyl)propanamide (50mg, 0.13 mmol) and methyl 2-(bromomethyl)benzoate (0.034 g, 0.15 mmol)was dissolved in acetonitrile (10 ml) and potassium carbonate (37 mg,0.27 mmol) was added. The mixture was stirred at 60° C. for three hours.Polymer supported trisamine (0.3 eqv) was added and stirred overnight.The polymer was filtered off and solvent was removed by evaporation.EtOAc (10 ml) was added and the organic phase was washed with threeportions of water. After drying the crude (MgSO₄), the solvent had beenremoved by evaporation. The residue was purified by preparative HPLC(started with acetonitrile/buffer 60/40 and then the acetonitrileconcentration was increased to 100%, the buffer was a mixture ofacetonitrile/water 10/90 and ammonium acetate (0.1 M, columnKR-100-7-C8, 50 mm×250 mm, flow 40 ml/min). The product containingfractions were pooled and the acetonitrile was removed by evaporation.EtOAc (10 ml) was added and the organic phase was washed with twoportions of brine and dried (MgSO₄). Removing the solvent by evaporationgave 15 mg of methyl2-[(4-{3-[butyl(2,3-dimethoxybenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoate(21.4%).

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.89-0.95 (m, 3H), 1.24-1.35 (m,2H), 1.48-1.57 (m, 2H), 2.62, 2.69 (t, t, 2H), 2.95, 3.01 (t, t, 2H),3.17, 3.36 (t, t, 2H), 3.83, 3.86, 3.89, 3.90, 3.93, 3.94 (s, s, s, s,s, s, 9H), 4.48, 4.71 (s, s, 2H), 5.50, 5.52 (s, s, 2H), 6.62, 6.75 (d,d, 1H), 6.84-6.97 (m, 3H), 7.00-7.04 (m, 1H), 7.11, 7.19 (d, d, 2H),7.38-7.42 (m, 1H), 7.56-7.60 (m, 1H), 7.78 (t, 1H) and 8.04-8.07 (m,1H).

c) Methyl2-[(4-{3-[butyl(2,3-dimethoxybenzyl)amino]-3-oxopropyl}phenoxy)methyl]-benzoate(15 mg, 0.029 mmol) was dissolved in THF/water (2/1, 2 ml) and LiOH (1.4mg, 0.058 mmol) was added. The reaction was performed in a single nodemicrowave oven (150° C., 7 min). Workup was done by adding EtOAc (10 ml)and washing the organic phase with two portions of HCl (2×5 ml, 1 M).The organic phase was dried (MgSO₄) and the solvent was removed byevaporation to give 13 mg of2-[(4-{3-[butyl(2,3-dimethoxybenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid (yield 89%).

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.89-0.95 (m, 3H), 1.24-1.35 (m,2H), 1.48-1.57 (m, 2H), 2.62, 2.69 (t, t, 2H), 2.95, 3.01 (t, t, 2H),3.17, 3.36 (t, t, 2H), 3.83, 3.85, 3.88, 3.89 (s, s, s, s, 6H), 4.48,4.71 (s, s, 2H), 5.50, 5.52 (s, s, 2H), 6.62, 6.75 (d, d, 1H), 6.84-6.97(m, 3H), 7.00-7.04 (m, 1H), 7.11, 7.19 (d, d, 2H), 7.38-7.42 (m, 1H),7.56-7.60 (m, 1H), 7.78 (t, 1H) and 8.04-8.07 (m, 1H).

¹³C NMR (rotamers, 125 MHz, CDCl₃): δ 13.76, 13.85, 20.02, 20.21, 29.55,30.30, 30.69, 30.84, 30.96, 35.13, 35.34, 42.60, 46.17, 46.37, 47.20,55.69, 55.75, 60.35, 61.74, 68.20, 111.23, 111.79, 114.88, 114.96,118.79, 120.88, 124.15, 124.21, 126.81, 127.18, 127.26, 129.46, 130.51,131.25, 131.52, 133.23, 133.65, 140.59, 146.50, 147.17, 152.48, 152.61,157.02, 157.10, 170.79, 172.93 and 173.25.

Example 8

a) N-(2,3-Dimethoxybenzyl)-N-heptylamine (0.70 g, 2.65 mmol) wasdissolved in DM (10 ml), 3-(4-hydroxyphenyl)propanoic acid (0.4 g, 2.41mmol), was added and the mixture was cooled to 0° C.N-[(1H-1,2,3-benzotriazol-1-yloxy)(dimethylamino)methylene]-N-methylmethanaminiumtetrafluoroborate (0.85 g, 2.65 mmol) and diisopropylethylamine (0.65 g,5.05 mmol) was added. The mixture was allowed to warm to roomtemperature and stirred overnight. EtOAc (15 ml) was added and theorganic phase was washed with two portions of sodium hydrogencarbonate(aq, 10 ml). EtOAc was removed by evaporation and the crude was purifiedby preparative HPLC (started with acetonitrile/buffer 60/40 and then theacetonitrile concentration was increased to 100%, the buffer was amixture of acetonitrile/water 10/90 and ammonium acetate (0.1 M, columnKR-100-7-C8, 50 mm×250 mm, flow 40 ml/min). The product containingfractions were pooled and the acetonitrile was removed by evaporation.EtOAc (10 ml) was added and the organic phase was washed with twoportions of brine and dried (MgSO₄). The solvent was removed byevaporation and gave 0.98 gramN-(2,3-dimethoxybenzyl)-N-heptyl-3-(4-hydroxyphenyl)propanamide (yield70%).

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.85-0.89 (m, 3H), 1.20-1.30 (m,8H), 1.47-1.56 (m, 2H), 2.62, 2.67 (t, t, 2H), 2.89, 2.95 (t, t, 2H),3.14, 3.33 (t, t, 2H), 3.79, 3.80, 3.84, 3.85 (s, s, s, s, 6H), 4.45,4.69 (s, s, 2M), 6.58, 6.68 (d, d, 1H), 6.74-6.88 (m, 3H) and 6.96-7.05(m, 3H).

b) N-(2,3-dimethoxybenzyl)-N-heptyl-3-(4-hydroxyphenyl)propanamide 0.196g, 0.47 mmol) and methyl 2-(bromomethyl)benzoate (0.12 g, 0.52 mmol) wasdissolved in acetonitrile (10 ml) and potassiumcarbonate (131 mg, 0.95mmol) was added. The mixture was stirred at 60° C. for three hours.Polymersupproted trisamine (0.3 eqv) was added and stirred over night.The polymer was filtered of, solvent was removed by evaporation,addition of EtOAc (10 ml) and the organic phase was washed with threepotions of water. After drying the crude (MgSO₄) and the solvent hadbeen removed by evaporation, the crude was purified by preparative HPLC(started with acetonitrile/buffer 60/40 and then the acetonitrileconcentration was increased to 100%, the buffer was a mixture ofacetonitrile/water 10/90 and ammonium acetate (0.1 M, columnKR-100-7-C8, 50 mm×250 mm, flow 40 ml/min). The product containingfractions was pooled and the acetonitrile was removed by evaporation.EtOAc (10 ml) was added and the organic phase was washed with twopotions of brine and dried (MgSO₄). Removing the solvent by evaporationgave 39 mg of methyl2-[(4-{3-[(2,3-dimethoxybenzyl)(heptyl)amino]-3-oxopropyl}phenoxy)methyl]benzoate(yield 14.6%).

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.86-0.89 (m, 3H), 1.19-1.30 (m,8H); 1.46-1.55 (m, 2H), 2.60, 2.66 (t, t, 2H), 2.93, 2.98 (t, t, 2H),3.14, 3.33 (t, t, 2H), 3.80, 3.83, 3.86, 3.87, 3.90, 3.91 (s, s, s, s,s, s, 9H), 4.45, 4.68 (s, s, 2H), 5.48, 5.49 (s, s, 2H), 6.59, 6.73 (d,d, 1H), 6.81-7.01 (m, 4H) 7.08, 7.16 (d, d, 2H) 7.35-7.39 (m, 1H),7.53-7.57 (m, 1H), 7.76 (t, 1H) and 8.01-8.04 (m, 1H).

c) Methyl2-[(4-{3-[(2,3-dimethoxybenzyl)(heptyl)amino]-3-oxopropyl}phenoxy)methyl]-benzoate(39 mg, 0.069 mmol) was dissolved in THF/water (2/1, 2 ml) and LiOH (3.3mg, 0.14 mmol) was added. The reaction was performed I a single nodemicrowave oven (150° C., 7 min). Workup was done by adding EtOAc (10 ml)and washing the organic phase with two potions of HCl (2×5 ml, 1 M). Theorganic phase was dried (MgSO₄) and the solvent was removed byevaporation to give 30 mg of2-[(4-{3-[(2,3-dimethoxybenzyl)(heptyl)-amino]-3-oxopropyl}phenoxy)methyl]benzoicacid (yield 78.9).

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.84-0.87 (m, 3H), 1.20-1.28 (m,8H), 1.45-1.55 (m, 2H), 2.63, 2.69 (t, t, 2H), 2.93, 2.99 (t, t, 2H),3.13, 3.33 (t, t, 2H), 3.79, 3.81, 3.84, 3.85 (s, s, s, s, 6H), 4.45,4.69 (s, s, 2H), 5.52, 5.53 (s, s, 2H), 6.60, 6.72 (d, d, 1H), 6.79-7.01(m, 4H) 7.08, 7.16 (d, d, 2H) 7.36-7.41 (m, 1H), 7.55-7.60 (m, 1H), 7.79(t, 1H) and 8.13-8.16 (m, 1H).

¹³C NMR (rotamers. 125 MHz. CDCl₃): δ 14.04, 22.52, 22.56, 26.77, 26.96,27.45, 28.60, 28.92, 29.03, 30.83, 30.97, 31.66, 31.75, 35.16, 35.36,42.61, 46.42,46.46,47.45, 55.69, 55.75, 60.35, 60.73, 68.19, 111.26,111.83, 114.88, 114.95, 118.84, 120.91, 124.14, 124.20, 126.85, 127.15,127.22, 129.46, 130.52, 131.26, 131.51, 133.19, 133.66, 140.61, 146.59,147.19, 152.48, 152.60, 157.02, 157.11, 170.77, 172.92 and 173.23.

Example 9 a) N-(3-ethoxypropyl)-N-(4-isopropylbenzyl)amine

p-iso-Propylbenzaldehyde (1.007 g, 6.798 mmol) was dissolved in methanol(5 ml). Trimethyl orthoformate (5 ml) was added. 3-Ethoxypropylamine(681 mg, 6.6 mmol) was then added and followed by acetic acid (0.2 ml).After standing at room temperature overnight, DCM (5 ml) was added andfollowed by borohydride on polymer support (5.28 g, 13.2 mmol). Themixture was shaken at room temperature for 4 days and then filtered. Thefiltrate was evaporated. The residue was dissolved in acetonitrile, thendivided into two portions and loaded on 2 columns (ISOLUTE® PRS, 10 g/70ml, wetted with acetonitrile). It was eluted with acetonitrile, thenmethanol and then methanol (NH₃ sat.). The product fractions werecombined and evaporated. Oil product 1.283 g was obtained, yield 83%.

¹H NMR (300 MHz, CDCl₃): δ 1.19 (t, 3H), 1.25 (d, 6H), 1.75-1.84 (m,2H), 2.73 (t, 2H), 2.85-2.94 (m, 1H), 3.43-3.52 (m, 4H), 3.75 (s, 2H),7.18 (d, 2H) and 7.24 (d, 2H).

b) N-(3-ethoxypropyl)-N-(4-isopropylbenzyl)amine (0.62 g, 2.65 mmol) wasdissolved in DMF (10 ml), 3-(4-hydroxyphenyl)propanoic acid (0.4 g, 2.41mmol), was added and the mixture was cooled to 0° C.N-[(1H-1,2,3-benzotriazol-1-yloxy)(dimethylamino)-methylene]-N-methylmethanaminiumtetrafluoroborate (0.85 g, 2.65 mmol) and diisopropylethylamine (0.65 g,5.05 mmol) was added. The mixture was allowed to warm to roomtemperature and stirred overnight. EtOAc (15 ml) was added and theorganic phase was washed with two portions of sodium hydrogencarbonate(aq, 10 ml). EtOAc was removed by evaporation and the crude was purifiedby preparative HPLC (started with acetonitrile/buffer 60/40 and then theacetonitrile concentration was increased to 100%, the buffer was amixture of acetonitrile/water 10/90 and ammonium acetate (0.1 M, columnKR-100-7-C8, 50 mm×250 mm, flow 40 ml/min). The product containingfractions were pooled and the acetonitrile was removed by evaporation.EtOAc (10 ml) was added and the organic phase was washed with twoportions of brine and dried (MgSO₄). The solvent was removed byevaporation and gave 1.0 gram ofN-(3-ethoxypropyl)-3-(4-hydroxyphenyl)-N-(4-isopropylbenzyl)propanamide(yield 75.8%).

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 1.16-1.21 (m, 3H), 1.26, 1.27 (d,d, 6H), 1.75-1.80, 1.84-1.90 (m, m, 2H), 2.64, 2.74 (t, t, 2H),2.86-3.00 (m, 3H), 3.33, 3.37 (t, t, 2H), 3.41-3.50 (m, 4H), 4.43, 4.62(s, s, 2H), 6.80-6.84 (m, 2H) and 6.97-7.22 (m, 6H).

c) N-(3-ethoxypropyl)-3-(4-hydroxyphenyl)-N-(4isopropylbenzyl)propanamide (0.18 g, 0.47 mmol) and methyl2-(bromomethyl)benzoate (0.12 g, 0.52 mmol) was dissolved inacetonitrile (10 ml) and potassium carbonate (131 mg, 0.95 mmol) wasadded. The mixture was stirred at 60° C. for three hours.Polymer-supported trisamine (0.3 eqv) was added and stirred overnight.The polymer was filtered off, solvent was removed by evaporation,addition of EtOAc (10 ml) and the organic phase was washed with threeportions of water. After drying the crude (MgSO₄) and the solvent hadbeen removed by evaporation, the crude was purified by preparative HPLC(started with acetonitrile/buffer 60/40 and then the acetonitrileconcentration was increased to 100%, the buffer was a mixture ofacetonitrile/water 10/90 and ammonium acetate (0.1 M, columnKR-100-7-C8, 50 mm×250 mm, flow 40 ml/min). The product containingfractions were pooled and the acetonitrile was removed by evaporation.EtOAc (10 ml) was added and the organic phase was washed with twopotions of brine and dried (MgSO₄). Removing the solvent by evaporationgave 0.16 gram of methyl2-[(4-{3-[(3-ethoxypropyl)(4-isopropylbenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoate(yield 63.5%).

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 1.17, 1.21 (t, t, 3H), 1.27, 1.28(d, d, 6H), 1.75-1.80, 1.84-1.89 (m, m, 2H), 2.63, 2.73 (t, t, 2H),2.89-3.04 (m, 3H), 3.32, 3.37 (t, t, 2H), 3.41-3.50 (m, 4H), 3.93, 3.94(s, s, 3H), 4.46, 4.61 (s, s, 2H), 5.51, 5.53 (s, s, 2H), 6.92, 6.95 (d,d, 2H), 7.04-7.22 (m, 6H), 7.40 (t, 1H), 7.58 (t, 1H), 7.78-7.80 (m, 1H)and 8.05-8.07 (m, 1H).

d) Methyl2-[(4-{3-[(3-ethoxypropyl)(4-isopropylbenzyl)amino]-3-oxopropyl}phenoxy)-methyl]benzoate(0.16 g, 0.30 mmol) was dissolved in THF/water (2/1, 2 ml) and LiOH(14.4 mg, 0.60 mmol) was added. The reaction was performed in a singlenode microwave oven (150° C., 7 min). Workup was done by adding EtOAc(10 ml) and washing the organic phase with two portions of HCl (2×5 ml,1 M). The organic phase was dried (MgSO₄) and the solvent was removed byevaporation to give 0.108 gram of2-[(4-{3-[(3-ethoxypropyl)(4-isopropylbenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid (yield 69.3%).

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 1.15, 1.19 (t, t, 3H), 1.23-1.25(m, 6H), 1.74-1.79, 1.84-1.89 (m, m, 2H), 2.66, 2.76 (t, t, 2H),2.86-3.03 (m, 3H), 3.30, 3.36 (t, t, 2H), 3.40-3.50 (m, 4H), 4.44, 4.61(s, s, 2H), 5.55, 5.56 (s, s, 2H), 6.90-6.94 (m, 2H), 7.02-7.20 (m, 6H),7.40 (t, 1H), 7.57-7.60 (m, 1H), 7.79-7.82 (m, 1H) and 8.15-8.18 (m,1H).

¹³C NMR (rotamers, 125 MHz, CDCl₃): δ 15.15, 23.98, 27.78, 28.61, 30.93,31.01, 33.73, 35.07, 35.38, 43.86, 44.13, 47.96, 51.26, 66.06, 66.22,66.82, 67.98, 68.13, 114.77, 126.03, 126.38, 126.72, 126.97, 127.90,129.31, 131.31, 132.88, 133.23, 133.39, 133.84, 134.56, 140.30, 140.38,147.71, 148.05, 156.90, 170.48, 173.02 and 173.26.

Example 10 a) N-(2,4-difluorobenzyl)-N-propylamine

2,4-Difluorbenzaldehyde (1.003 g, 7.055 mmol) was dissolved in methanol(5 ml). Trimethyl orthoformate (5 ml) was added. Propylamine (401 mg,6.784 mmol) was then added and followed by acetic acid (0.2 ml). After 1hour, DCM (5 ml) was added and followed by borohydride polymer-supported(2.5 mmol/g, 5.42 g, 13.55 mmol). The mixture was shaken at roomtemperature for 4 days and then filtered. The filtrate was evaporated.The residue was dissolved in acetonitrile, then divided into twoportions and loaded on two columns (ISOLUTE®PRS, 10 g/70 ml, wetted withacetonitrile). It was eluted with acetonitrile, then methanol and thenmethanol (NH₃ sat.). The product fractions were combined and evaporated.Oil product (892 mg) was obtained, yield 71%.

¹H NMR (400 MHz, CDCl₃): δ 0.91 (t, 3H), 1.47-1.56 (m, 2H), 2.57 (t,2H), 3.79 (s, 2H), 6.75-6.85 (m, 2H) and 7.27-7.33 (m, 1H).

b) N-(2,4-difluorobenzyl)-3-(4-hydroxyphenyl)-N-propylpropanamide

3-(4-Hydroxyphenyl)propionic acid (245 mg, 1.474 mmol) in DMF (5 ml) wascooled in an ice-bath. N-(2,4-Difluorobenzyl)-N-propylamine (300.4 mg,1.622 mmol) was added and then TBTU (521 mg, 1.622 mmol) followed byDIPEA (400 mg, 3.096 mmol). The mixture was stirred at room temperatureovernight. Sodium hydrogencarbonate aqueous solution (sat.) was added.The mixture was extracted with ethyl acetate (x2). The extracts werecombined and dried (magnesium sulphate) and evaporated. Chromatographyof the residue on a column (ISOLUTE® SI, 5 g/25 ml) using DCM/heptane(50:50), DCM and then MeOH/DCM (1:99, then 2:98) as eluant gave 336 mgthe desired product, yield 68%.

¹H NMR (rotamers, 300 MHz, CDCl₃): δ 0.82-0.88 (m, 3H), 1.45-1.58 (m,2H), 2.59, 2.65 (t, t, 2H), 2.88-2.98 (m, 2H), 3.11, 3.27 (t, t, 2H),4.40, 4.59 (s, s, 2H), 6.71-7.03 (m, 6H), 7.07-7.16 (m, 1H) and 7.79 (s,br, 1H).

c) Methyl2-[(4-{3-[(2,4-difluorobenzyl)(propyl)amino]-3-oxopropyl}phenoxy)methyl]benzoate

N-(2,4-Difluorobenzyl)-3-(4-hydroxyphenyl)-N-propylpropanamide (290 mg,0.87 mmol) was dissolved in acetonitrile (10 ml). 2-Bromomethyl-benzoicacid methyl ester (209 mg, 0.913 mmol) was added followed by potassiumcarbonate, anhydrous (180 mg, 1.305 mmol). The mixture was heated toreflux overnight and then evaporated to dry. Water and ethyl acetatewere added and two phases were separated. The organic phase was dried(magnesium sulphate) and evaporated. Chromatography of the residue on acolumn (ISOLUTE®SI, 20 g/70 ml) using DCM and then MeOH/DCM (1:99) aseluant gave 184 mg the desired product, yield 44%.

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 0.85-0.92 (m, 3H), 1.52-1.60 (m,2H), 2.61, 2.68 (t, t, 2H), 2.95-3.01 (m, 2H), 3.15, 3.32 (t, t, 2H),3.92, 3.93 (s, s, 3H), 4.45, 4.62 (s, s, 2H), 5.51, 5.52 (s, s, 2H),6.77-6.86 (m, 2H), 6.92-6.99, 7.23-7.27 (m, m, 3H), 7.12, 7.16 (d, d,2H), 7.38-7.42 (m, 1H), 7.57 (t, 1H) 7.78 (d, 1H) and 8.04-8.07 (m, 1H).

d)2-[(4-{3-[(2,4-difluorobenzyl)(propyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid

A mixture of methyl2-[(4-{3-[(2,4-difluorobenzyl)(propyl)amino]-3-oxopropyl}phenoxy)-methyl]benzoate(0.184 g, 0.382 mmol) and lithium hydroxide (0.018 g, 0.76 mmol) in THF(2 ml) and water (2 ml) was heated at 150 degrees for 7 minutes. Themixture was diluted with water, acidified with hydrochloric acid andextracted with methylene chloride. The combined extracts were dried withmagnesium sulfate and evaporated to give2-[(4-{3-[(2,4-difluorobenzyl)(propyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid.

¹HNMR (400 MHz, CDCl₃): δ 0.85 (t, 3), 1.4-1.6 (m, 2), 2.6-2.7 (m, 2),2.9-3.0 (m, 2), 3.05-3.15 and 3.25-3.35 (multiplets, rotamers, 2), 4.4and 4.6 (singlets, rotamers, 2), 5.5 (m, 2), 6.7-6.8 (m, 2), 6.9-7.0 (m,2), 7.05-7.2 (m, 2), 7.4 (t, 1), 7.6 (t, 1), 7.8 (d, 1), 8.15 (d, 1).

Example 11

a) Methyl2-({[4-(2-{ethyl[4-(trifluoromethyl)benzyl]amino}-2-oxoethyl)phenyl]thio}methyl)benzoate

Ethyl[4-(trifluoromethyl)benzyl]amine (284 mg, 1.40 mmol) and(4-{[2-(methoxycarbonyl)benzyl]thio}phenyl)acetic acid (443 mg, 1.40mmol, See Example 2 b) were mixed in DMF (15 ml). TBTU (472 mg, 1.47mmol) was added and then DIPEA (190 mg, 1.47 mmol) was added. Themixture was stirred at room temperature for 1 hour. EtOAc (20 ml) wasadded. The mixture was then washed with sodium hydrogencarbonate aqueoussolution (sat.), 1% HCl, water (x2) and brine, dried with magnesiumsulphate and evaporated. The title compound (663 mg) was left, yield94%.

¹H NMR (rotamers, 500 MHz, CDCl₃): δ 1.08-1.15 (m, 3H), 3.31, 3.45 (q,q, 2H), 3.63, 3.76 (s, s, 2H), 3.92 (s, 3H), 4.524.66 (m, 4H), 7.09-7.40(m, 8H), 7.56, 7.62 (d, d, 2H) and 7.94 (d, 2H).

b)2-({[4-(2-{Ethyl[4-(trifluoromethyl)benzyl]amino}-2-oxoethyl)phenyl]thio}methyl)benzoicacid

Methyl2-({[4-(2-{ethyl[4-(trifluoromethyl)benzyl]amino}-2-oxoethyl)phenyl]thio}methyl)benzoate(640 mg, 1.276 mmol) was dissolved in THF (20 ml). Lithium hydroxide (61mg, 2.552 mmol) in water (10 ml) was added. The mixture was stirred atroom temperature and the reaction was followed by LC-MS. After 24 hours,more lithium hydroxide (30 mg) and water (10 ml) were added. After intotal 70 hour, LC-MS showed the reaction was complete. The mixture wasevaporated to remove THF. The residue was washed with diethyl ether,then acidified with 10% HCl, pH=2, and extracted with EtOAc (x2). Theextracts were combined and washed with brine, dried with magnesiumsulphate and evaporated. Chromatography of the residue on a column(ISOLUTE SI, 5 g/25 ml), using DCM and then MeOH/DCM (0.5:99.5, then1:99) as eluant, gave the title compound (510 mg), yield 82%.

¹H NMR (rotamers, 300 MHz, CDCl₃): δ 1.07-1.15 (m, 3H), 3.31, 3.45 (q,q, 2H), 3.64, 3.77 (s, s, 2H), 4.54 (s, 2H), 4.55, 4.66 (s, s, 2H),7.05-7.40 (m, 8H), 7.55, 7.60 (d, d, 2H) and 7.97 (d, 2H).

¹³C NMR (rotamers, 75 MHz, CDCl₃): δ 12.60, 13.80, 38.17, 38.25, 40.00,40.44, 41.63, 42.40, 47.94, 50.61, 118-134 (complex multiplet), 140.32,140.70, 141.50, 170.48 and 171.20.

Example 12

2-[(4-{2-[Ethyl(2-fluorobenzyl)amino]-2-oxoethyl}phenoxy)methyl]benzoicacid.

a) N-(2-Fluorobenzyl)ethanamine (0.554 g, 3.615 mmol) was dissolved inDMF (10 ml). (4-Hydroxyphenyl)acetic acid (0.500 g, 3.286) was added andthe mixture was cooled to 0° C.N-[(1H-1,2,3-Benzotriazol-1-yloxy)(dimethylamino)methylene]-N-methylmethanaminiumtetrafluoroborate (1.161 g, 3.615 mmol) and N-ethyl-N,N-diisopropylamine(0.892 g, 6.901 mmol) were added. The solution was stirred overnight atroom temperature. EtOAc (20 ml) was added and the organic phase waswashed with two portions of Na₂CO3 (2×20 ml, aq). The organic layer wasdried (MgSO₄) and the solvent was removed by evaporation. The crude waspurified by preparative HPLC (started with isocratic acetonitrile/buffer60/40 and then the acetonitrile concentration was increased to 100%, thebuffer was a mixture of acetonitrile/water 10/90 and ammonium acetate(0.1 M, column KR-100-7-C8, 50 mm×250 mm, flow 40 ml/min). Theproduct-containing fractions were pooled and the acetonitrile wasremoved by evaporation. EtOAc (10 ml) was added and the organic phasewas washed with two portions of brine and dried (MgSO₄) and the solventwas removed by evaporation to give 0.69 g ofN-ethyl-N-(2-fluorobenzyl)-2-(4-hydroxyphenyl)acetamide (yield 73.1%).

¹HNMR (Rotamers, 500 MHz, CDCl₃): δ 1.12 (m, 3H), 3.35-3.5 (m, 2H),3.62-3.72 (m, 2H), 4.57-4.78 (m, 2H), 6.7 (t, 2H), 6.98-7.38 (m, 6H).

b) N-Ethyl-N-(2-fluorobenzyl)-2-(4-hydroxyphenyl)acetamide (0.381 g,1.327 mmol) and methyl 2-(bromomethyl)benzoate (0.334 g, 1.460 mmol)were dissolved in acetonitrile (10 ml) and potassium carbonate (0.367 g,2.654 mmol) was added. The mixture was stirred at 65° C. for threehours. When N-ethyl-N-(2-fluorobenzyl)-2-(4-hydroxyphenyl)acetamide wasconsumed, PS-trisamine (0.3 eqv) was added and the solution was stirredovernight at room temperature. The polymer was filtered off andacetonitrile was removed by evaporation. EtOAc (10 ml) was added and theorganic phase was washed with two portions of water, dried (MgSO₄) andthe solvent was removed by evaporation to give 0.545 g of methyl2-[(4-{2-[ethyl(2-fluorobenzyl)amino]-2-oxoethyl}phenoxy)methyl]benzoate(yield 94.3%).

¹HNMR (Rotamers, 300 MHz, CDCl₃): δ 1.12 (m, 3H), 3.23-3.35 (m, 2H),3.6-3.75 (m, 2H), 3.88 (s, 3H), 4.45-4.70 (m, 2H), 5.45 (s, 2H),6.84-7.35 (m, 9H), 7.5 (t, 1H), 7.72 (d, 1H), 8.0 (d, 1H).

c) Methyl2-[(4-{2-[ethyl(2-fluorobenzyl)amino]-2-oxoethyl}phenoxy)methyl]-benzoate(0.545 g, 1.251 mmol) was dissolved in EtOH (5 ml) and potassiumhydroxide (0.105 g, 1.877 mmol) was added. The reaction was performed ina single node microwave oven (7 min, 150° C.). Workup was by removingthe solvent by evaporation, addition of HCl (20 ml, 1 M) and the waterphase was washed with two portions of EtOAc (20 ml). The organic phaseswere pooled and the solvent was removed by evaporation. The crude waspurified by preparative HPLC (starting with isocraticacetonitrile/buffer 60/40 and then the acetonitrile concentration wasincreased to 100%, the buffer was a mixture of acetonitrile/water 10/90and ammonium acetate (0.1 M, column KR-100-7-C8, 50 mm×250 mm, flow 40ml/min). The product-containing fractions were pooled and theacetonitrile was removed by evaporation. EtOAc (10 ml) was added and theorganic phase was washed with two portions of brine and dried (MgSO₄).The solvent was removed by evaporation to give 0.124 g of2-[(4-{2-[ethyl(2-fluorobenzyl)amino]-2-oxoethyl}phenoxy)methyl]benzoicacid (23.5%).

¹HNMR (Rotamers, 500 MHz, CDCl₃): δ 1.12 (m, 3H), 3.25-3.5 (m, 2H),3.65-3.8 (m, 2H), 4.5-4.75 (m, 2H), 5.52 (m, 2H), 6.84-7.45 (m, 9H),7.55 (t, 1H), 7.78 (d, 1H), 8.13 (d, 1H).

Example 13

2-[(4-{3-[Ethyl(2-fluorobenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid.

a) N-(2-Fluorobenzyl)ethanamine (0.554 g, 3.615 mmol) was dissolved inDMF (10 ml). 3-(4-hydroxyphenyl)propanoic acid (0.546 g, 3.286 mmol) wasadded and the mixture was cooled to 0° C.N-[(1H-1,2,3-benzotriazol-1-yloxy)(dimethylamino)-methylene]-N-methylmethanaminiumtetrafluoroborate (1.161 g, 3.615 mmol) and N-ethyl-N,N-diisopropylamine(0.892 g, 6.901 mmol) was added. The solution was stirred overnight atroom temperature. EtOAc (20 ml) was added and the organic phase waswashed with two portions of Na₂CO₃ (2×20 ml, aq). The organic layer wasdried (MgSO₄) and the solvent was removed by evaporation. The crude waspurified by preparative HPLC (starting with isocraticacetonitrile/buffer 60/40 and then the acetonitrile concentration wasincreased to 100%, the buffer was a mixture of acetonitrile/water 10/90and ammonium acetate (0.1 M, column KR-100-7-C8, 50 mm×250 mm, flow 40ml/min). The product-containing fractions were pooled and theacetonitrile was removed by evaporation. EtOAc (10 ml) was added and theorganic phase was washed with two portions of brine and dried (MgSO4)and then solvent was removed by evaporation to give 0.803 g ofN-ethyl-N-(2-fluorobenzyl)-3-(4-hydroxyphenyl)propanamide (yield 81.1%).

¹HNMR (Rotamers, 500 M, CDCl₃): δ 1.1 (m, 3H), 2.58-2.72 (m, 2H),2.83-3.0 (m, 2H), 3.2-3.5 (m, 2H), 4.45-4.7 (m, 2H), 6.78 (t, 2H),6.95-7.35 (m, 6H).

b) N-Ethyl-N-(2-fluorobenzyl)-3-(4-hydroxyphenyl)propanamide (0.400 g,1.327 mmol) and methyl 2-(bromomethyl)benzoate (0.334 g, 1.460 mmol)were dissolved in acetonitrile (10 ml) and potassium carbonate (0.367 g,2.654 mmol) was added. The mixture was stirred at 65° C. for threehours. When N-ethyl-N-(2-fluorobenzyl)-2-(4-hydroxyphenyl)acetamide wasconsumed, PS-trisamine (0.3 eqv) was added and the solution was stirredovernight at room temperature. The polymer was filtered off andacetonitrile was removed by evaporation. EtOAc (10 ml) was added and theorganic phase was washed with two portions of water, dried (MgSO₄) andthe solvent was removed by evaporation. EtOAc (10 ml) was added and theorganic phase was washed with two portions on water, dried (MgSO₄) andthe solvent was removed by evaporation. The crude was purified bypreparative HPLC (starting with isocratic acetonitrile/buffer 60/40 andthen the acetonitrile concentration was increased to 100%, the bufferwas a mixture of acetonitrile/water 10/90 and ammonium acetate (0.1 M,column KR-100-7-C8, 50 mm×250 mm, flow 40 ml/min). Theproduct-containing fractions were pooled and the acetonitrile wasremoved by evaporation. EtOAc (10 ml) was added and the organic phasewas washed with two portions of brine and dried (MgSO₄). The solvent wasremoved by evaporation.to give 0.454 g of methyl2-[(4-{3-[ethyl(2-fluorobenzyl)amino]-3-oxopropyl}phenoxy)methyl]-benzoate(yield 76.1%).

¹HNMR (Rotamers, 400 MHz, CDCl₃): δ 1.08 (m, 3H), 2.52-2.68 (m, 2H),2.9-3.03 (m, 2H), 3.18-3.45 (m, 2H), 3.88 (s, 3H), 4.454.7 (m, 2H), 5.45(s, 2H), 6.82-7.28 (m, 8H), 7.36 (t, 1H), 7.53 (t, 1H), 7.73 (d, 1H),8.0 (d, 1H).

c) Methyl2-[(4-{3-[ethyl(2-fluorobenzyl)amino]-3-oxopropyl}phenoxy)methyl]-benzoate(0.454 g, 1.001 mmol) was dissolved in EtOH (5 ml) and potassiumhydroxide (0.085 g, 1.514 mmol) was added. The reaction was performed ina single node microwave oven (7 min, 150° C.). Workup was by removingthe solvent by evaporation, addition of HCl (20 ml, 1 M) and the waterphase was washed with two portions of EtOAc (20 ml). The organic phaseswere pooled and the solvent was removed by evaporation. The crude waspurified by preparative HPLC (starting with isocraticacetonitrile/buffer 60/40 and then the acetonitrile concentration wasincreased to 100%, the buffer was a mixture of acetonitrile/water 10/90and ammonium acetate (0.1 M, column KR-100-7-C8, 50 mm×250 mm, flow 40ml/min). The product-containing fractions were pooled and theacetonitrile was removed by evaporation. EtOAc (10 ml) was added and theorganic phase was washed with two portions of brine and dried (MgSO₄).The solvent was removed by evaporation to give 0.079 g of2-[(4-{3-[ethyl(2-fluorobenzyl)amino]-3-oxopropyl}-phenoxy)methyl]benzoicacid (18.0%).

¹HNMR (Rotamers, 500 MHz, CDCl₃): δ 1.12 (m, 3H), 2.60-2.75 (m, 2H),3.03 (m, 2H), 3.23-3.50 (m, 2H), 4.45-4.73 (m, 2H), 5.57 (s, 2H),6.88-7.35 (m, 8H), 7.41 (t, 1H), 7.60 (t, 1H), 7.80 (d, 1H), 8.18 (d,1H).

The following Examples were prepared in a similar manner.

Example 14

2-{[(3-{2-[(2,4-Difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoicacid.

Example 15

2-{[(4-{2-[(4-Chlorobenzyl)(ethyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoicacid.

Biological Activity

Formulations

Compounds were dissolved in DMSO to obtain 16 mM stock solutions. Beforeassays, stock solutions were further diluted in DMSO and culture media.

General Chemicals and Reagents

Luciferase assay reagent was purchased from Packard, USA. RestrictionEnzymes were from Boehringer and Vent polymerase from New EnglandBiolabs.

Cell Lines and Cell Culture Conditions

U2-OS, (Osteogenic sarcoma, Human) was purchased from ATCC, USA. Cellswere expanded and refrozen in batches from passage number six. Cellswere cultured in Dulbecco's modified Eagle medium (DMEM) with 25 mMglucose, 2 mM glutamine or 4 mM L-alanyl-L-glutamine, 10% fetal calfserum, at 5% CO₂. Phosphate buffered saline (PBS) without addition ofcalcium or magnesium was used. All cell culture reagents were from Gibco(USA) and 96-well cell culture plates were purchased from Wallach.

Plasmid Constructs for Heterologous Expression

Standard recombinant DNA techniques were carried out as described byAusubel (7). The Luciferase reporter vector, pGL5UAS (clone consists offive copies of the GAL4 DNA binding sequence,5′-CGACGGAGTACTGTCCTCCGAGCT-3′, cloned into the SacI/XhoI sites ofpGL3-Promoter (Promega). The SacI/XhoI fragment carrying the UAS siteswas constructed using annealed overlapping oligonucleotides.

Expression vectors used are based upon pSG5 (Stratagene). All vectorscontain an EcoRI/NheI fragment encoding the DNA binding domain of GAL4(encoding amino acid positions 1-145 of database accession numberP04386) followed by an in-frame fusion to a fragment encoding thenuclear localisation sequence from T antigen of Polyoma Virus. Thenuclear localisation sequence was constructed using annealed overlappingoligonucleotides creating NheI/KpnI sticky ends (5CTAGCGCTCCTAGAAGAAACGCAAGGTTGGTAC-3′). The ligand binding domains fromhuman and mouse PPARα and human and mouse PPARγ were PCR amplified asKpnI/BamHI fragments and cloned in frame to the GAL4 DNA binding domainand the nuclear localisation sequence. The sequence of all plasmidconstructs used were confirmed by sequencing.

The following expression vectors were used for transient transfections:vector encoded PPAR subtype sequence reference¹ pSGGALhPPa human PPARαS74349, nt 625-1530 pSGGALmPPa murine PPARα X57638, nt 668-1573pSGGALhPPg human PPARγ U63415, nt 613-1518 pSGGALmPPg murine PPARγU09138, nt 652-1577¹refers to nucleotide positions of data base entry used to express theligand binding domain.Transient Transfections

Frozen stocks of cells from passage number six were thawed and expandedto passage number eight before transfections. Confluent cells weretrypsinised, washed and pelleted by centrifugation at 270×g for 2minutes. The cell pellet was resuspended in cold PBS to a cellconcentration of about 18×10⁶ cells/ml. After addition of DNA, the cellsuspension was incubated on ice for approximately 5 minutes beforeelectroporation at 230 V, 960 μF in Biorad's Gene Pulser™ in 0.5 mlbatches. A total of 50 μg DNA was added to each batch of 0.5 ml cells,including 2.5 μg expression vector, 25 μg reporter vector and 22.5 μgunspecific DNA (pBluescript, Stratagene).

After electroporation, cells were diluted to a concentration of 320'000cells/ml in DMEM without phenol red, and approximately 25'000 cells/wellwere seeded in 96-well plates. In order to allow cells to recover,seeded plates were incubated at 37° C. for 3-4 hours before addition oftest compounds. In assays for PPARα, the cell medium was supplementedwith resin-charcoal stripped fetal calf serum (FCS) in order to avoidbackground activation by fatty acid components of the FCS. Theresin-charcoal stripped FCS was produced as follows; for 500 ml ofheat-inactivated FCS, 10 g charcoal and 25 g Bio-Rad Analytical GradeAnion Exchange Resin 200-400 mesh were added, and the solution was kepton a magnetic stirrer at room temperature over night. The following day,the FCS was centrifuged and the stripping procedure was repeated for 4-6hours. After the second treatment, the FCS was centrifuged and filtersterilised in order to remove remnants of charcoal and resin.

Assay Procedure

Stock solutions of compounds in DMSO were diluted in appropriateconcentration ranges in master plates. From master plates, compoundswere diluted in culture media to obtain test compound solutions forfinal doses.

After adjustment of the amount of cell medium to 75 μl in each well, 50μl test compound solution was added. Transiently transfected cells wereexposed to compounds for about 24 hours before the luciferase detectionassay was performed. For luciferase assays, 100 μl of assay reagent wasadded manually to each well and plates were left for approximately 20 tominutes in order to allow lysis of the cells. After lysis, luciferaseactivity was measured in a 1420 Multiwell counter, Victor, from Wallach.

Reference Compounds

The TZD pioglitazone was used as reference substance for activation ofboth human and murine PPARγ. 5,8,11,14-Eicosatetrayonic acid (ETYA) wasused as reference substance for human PPARα.

Calculations and Analysis

For calculation of EC₅₀ values, a concentration-effect curve wasestablished. Values used were derived from the average of two or threeindependent measurements (after subtraction of the background averagevalue) and were expressed as the percentage of the maximal activationobtained by the reference compound. Values were plotted against thelogarithm of the test compound concentration. EC₅₀ values were estimatedby linear intercalation between the data points and calculating theconcentration required to achieve 50% of the maximal activation obtainedby the reference compound.

The compounds of formula I have an EC₅₀ of less than 50 μmol/A for PPARαand preferred compounds have an EC₅₀ of less than 5 μmol/l. For examplethe EC₅₀s of some of the Examples for human PPAR alpha are: Example 50.163 μmol/l; Example 10 0.168 μmol/l; Example 11 0.026 μmol/l; andExample 15 0.027 μmol/l.

1. A compound of formula I

wherein n is 0, 1 or 2 and R¹ represents halo, a C₁₋₄alkyl group whichis optionally substituted by one or more fluoro, a C₁₋₄alkoxy groupwhich is optionally substituted by one or more fluoro and wherein when nis 2 the substituents R¹ may be the same or different; R² represents aC₂₋₈alkyl group which is optionally interrupted by oxygen; Y is absentor represents methylene; and X is O or S; and pharmaceuticallyacceptable salts and prodrugs thereof.
 2. A compound according to claim1 in which X is O.
 3. A compound according to claim 1 in which X is S.4. A compound according to any preceding claim in which Y is methylene.5. A compound according to any one of claims 1, 2 or 3 in which Y isabsent.
 6. A compound according to any preceding claim in which R¹ ishalo, a C₁₋₄alkyl group or a C₁₋₄alkoxy group and n is 0, 1 or
 2. 7. Acompound according to any preceding claim in which R¹ is fluoro,methoxy, or isopropyl when n is 1 or
 2. 8. A compound according to anypreceding claim in which n is
 0. 9. A compound according to anypreceding claim in which R² represents a C₅₋₇alkyl group.
 10. A compoundselected from:2-[(4-{3-[benzyl(hexyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic acid;2-{[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoicacid; 2-[(4-{2-[benzyl(hexyl)amino]-2-oxoethyl}phenoxy)methyl]benzoicacid;2-[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenoxy)methyl]benzoicacid;2-[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid;2-{[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenyl)thio]methyl}benzoicacid;2-[(4-{3-[butyl(2,3-dimethoxybenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid;2-[(4-{3-[(2,3-dimethoxybenzyl)(heptyl)-amino]-3-oxopropyl}phenoxy)methyl]benzoicacid;2-[(4-{3-[(3-ethoxypropyl)(4-isopropylbenzyl)amino]-3-oxopropyl}phenoxy)methyl]-benzoicacid;2-[(4-{3-[(2,4-difluorobenzyl)(propyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid;2-[(4-{2-[ethyl(2-fluorobenzyl)amino]-2-oxoethyl}phenoxy)methyl]benzoicacid;2-[(4-{3-[ethyl(2-fluorobenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid;2-({[4-(2-{ethyl[4-(trifluoromethyl)benzyl]amino}-2-oxoethyl)phenyl]thio}-methyl)benzoicacid;2-{[(3-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoicacid; and2-{[(4-{2-[(4-chlorobenzyl)(ethyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoicacid and pharmaceutically acceptable salts thereof.
 11. A compoundaccording to claim 2 selected from:2-[(4-{3-[benzyl(hexyl)amino]-3-oxopropyl}phenoxy)methyl]benzoic acid;2-[(4-{2-[benzyl(hexyl)amino]-2-oxoethyl}phenoxy)methyl]benzoic acid;2-[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenoxy)methyl]benzoicacid;2-[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid;2-[(4-{3-[butyl(2,3-dimethoxybenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid;2-[(4-{3-[(2,3-dimethoxybenzyl)(heptyl)-amino]-3-oxopropyl}phenoxy)methyl]benzoicacid;2-[(4-{3-[(3-ethoxypropyl)(4-isopropylbenzyl)amino]-3-oxopropyl}phenoxy)methyl]-benzoicacid;2-[(4-{3-[(2,4-difluorobenzyl)(propyl)amino]3-oxopropyl}phenoxy)methyl]benzoicacid;2-[(4-{2-[ethyl(2-fluorobenzyl)amino]-2-oxoethyl}phenoxy)methyl]benzoicacid;2-[(4-{3-[ethyl(2-fluorobenzyl)amino]-3-oxopropyl}phenoxy)methyl]benzoicacid; and pharmaceutically acceptable salts thereof.
 12. A compoundaccording to claim 3 selected from:2-{[(4-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoicacid;2-{[(4-{3-[(2,4-difluorobenzyl)(heptyl)amino]-3-oxopropyl}phenyl)thio]methyl}benzoicacid;2-({[4-(2-{ethyl[4-(trifluoromethyl)benzyl]amino}-2-oxoethyl)phenyl]thio}-methyl)benzoicacid;2-{[(3-{2-[(2,4-difluorobenzyl)(heptyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoicacid; and2-{[(4-{2-[(4-chlorobenzyl)(ethyl)amino]-2-oxoethyl}phenyl)thio]methyl}benzoicacid and pharmaceutically acceptable salts thereof.
 13. A pharmaceuticalformulation comprising a compound according to any preceding claim inadmixture with pharmaceutically acceptable adjuvants, diluents and/orcarriers.
 14. A method of treating or preventing insulin resistancecomprising the administration of a compound according to any one ofclaims 1 to 12 to a mammal in need thereof.
 15. The use of a compoundaccording to any one of claims 1 to 12 in the manufacture of amedicament for the treatment of insulin resistance.
 16. A process toprepare a compound of formula I comprising reacting a compound offormula II

in which R¹, R², X and Y are as previously defined and PG represents aprotecting group for a carboxylic hydroxy group with a de-protectingagent.
 17. A compound of formula II as described in claim
 16. 18. Acombination treatment comprising a compound according to any one ofclaims 1 to 12 in combination with another therapeutic agent that isuseful in the treatment of disorders associated with the development andprogress of atherosclerosis, hypertension, hyperlipidaemias,dyslipidaemias, diabetes and obesity.